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Volume 27,Issue 3,2020 Table of Contents
2020, 27(3):213-220.
DOI: 10.3872/j.issn.1007-385X.2020.03.001
Abstract:
New strategies for cancer immunotherapy have developed rapidly in recent years. Regarding how to achieve the optimal anti-tumor immune effect,“passive”immunotherapy such as adoptive cell transfer therapy, genetically engineered T cells, etc. can be used to directly attack tumor cells while“active”immunotherapy such as cytokines, cancer vaccines, immune checkpoint inhibitors,etc. can be used to regulate and activate immune system. In situ vaccine, using intratumoral administration of immunomodulators,combines“active”and“passive”immunotherapies scientifically at the same time, forming a cycle composed of immune priming-immune activition-tumor cell death-antigen release resulting in immune priming-immune activition, ultimately maximizing the anti-tumor immune effect. This review describes the specific strategies, promising preclinical results and some clinical trials as well as advantages,challenges and perspectives of in situ vaccine.
New strategies for cancer immunotherapy have developed rapidly in recent years. Regarding how to achieve the optimal anti-tumor immune effect,“passive”immunotherapy such as adoptive cell transfer therapy, genetically engineered T cells, etc. can be used to directly attack tumor cells while“active”immunotherapy such as cytokines, cancer vaccines, immune checkpoint inhibitors,etc. can be used to regulate and activate immune system. In situ vaccine, using intratumoral administration of immunomodulators,combines“active”and“passive”immunotherapies scientifically at the same time, forming a cycle composed of immune priming-immune activition-tumor cell death-antigen release resulting in immune priming-immune activition, ultimately maximizing the anti-tumor immune effect. This review describes the specific strategies, promising preclinical results and some clinical trials as well as advantages,challenges and perspectives of in situ vaccine.
SHI Long
,
GENG Songsong
,
CAI Ziqi
,
HAN Jinsheng
,
ZHAO Zhilong
,
ZHANGWei
,
SONG Hongtao
,
MENG Tongyu
,
CAI Jianhui
2020, 27(3):221-227.
DOI: 10.3872/j.issn.1007-385X.2020.03.002
Abstract:
Objective: To investigate the anti-tumor effect of CTL cells on colon cancer xenograft in nude mice after knocking out the immune check point CTLA-4 by CRISPR/Cas9 technology. Methods: A specific small guide RNA (sgRNA) for CTLA-4 was designed to construct sgRNA/Cas9 plasmid, which was then transfected into CTL using a lentiviral vector to obtain CTL cells with CTLA-4 deletion (CTLA-4 KO CTL). The transfection efficiency of the plasmid and the deletion efficiency of CTLA-4 were verified. BALB/c nude mice were randomly divided into two groups to prophylactically inoculate CTLA-4 KO CTL (experimental group) or CTL (control group); 3 days later, the animals of two groups were inoculated with colon cancer cell line LS174-T to observe the tumor formation rate and tumor formation time. After constructing colon cancer xenograft model in nude mice, the animals were randomly divided into two groups, respectively treated with CTLA-4 KO CTL (experimental group) and CTL (control group) cells to observe the tumor growth volume and survival time of mice. The serum levels of TNF-α and IFN-γ in nude mice were detected. Results: sgRNA was designed and CRSIPR/Cas9 system with lentivirus as vector was successfully constructed. CTL cells were transfected with the established CRSIPR/Cas9 system, and the highest transfection efficiency was up to (28.80±0.62)%. After transfection, the deletion efficiency of CTLA-4 was detected by Flow cytometry. The CTLA-4 expression of CTLA-4 KO CTL group was significantly lower than that of CTL group [(0.91±0.25)% vs (42.70±2.72)%, P<0.05]. In prophylactic assay, the formation rate of colon cancer xenografts in the experimental group was significantly lower than that in the control group (33.33%vs 100%, P<0.05). In treatment assay, the tumor volume in the experimental group was significantly inhibited compared with the control group ([503±23.9] vs [911.2±51.4] mm3, P<0.05), and the survival time of the experimental group was significantly prolonged (median survival time: 78 d vs 42 d, P<0.05); Moreover, the secretion levels of serumTNF-α ([268.93±17.04] pg/ml vs [148.26±20.07] pg/ml, P<0.05) and IFN-γ (315.38±18.67 pg/ml vs 202.92±29.32 pg/ml,P<0.05) in the experimental group were significantly higher than those in the control group. Conclusions: The lentiviral vector CRSIPR/Cas9 system is an effective gene editing method; its successful deletion of CTLA-4 in CTL cells can significantly inhibit the tumor formation rate of colon cancer xenografts in nude mice and enhance the anti-tumor effect of CTL on colon cancer xenografts.
Objective: To investigate the anti-tumor effect of CTL cells on colon cancer xenograft in nude mice after knocking out the immune check point CTLA-4 by CRISPR/Cas9 technology. Methods: A specific small guide RNA (sgRNA) for CTLA-4 was designed to construct sgRNA/Cas9 plasmid, which was then transfected into CTL using a lentiviral vector to obtain CTL cells with CTLA-4 deletion (CTLA-4 KO CTL). The transfection efficiency of the plasmid and the deletion efficiency of CTLA-4 were verified. BALB/c nude mice were randomly divided into two groups to prophylactically inoculate CTLA-4 KO CTL (experimental group) or CTL (control group); 3 days later, the animals of two groups were inoculated with colon cancer cell line LS174-T to observe the tumor formation rate and tumor formation time. After constructing colon cancer xenograft model in nude mice, the animals were randomly divided into two groups, respectively treated with CTLA-4 KO CTL (experimental group) and CTL (control group) cells to observe the tumor growth volume and survival time of mice. The serum levels of TNF-α and IFN-γ in nude mice were detected. Results: sgRNA was designed and CRSIPR/Cas9 system with lentivirus as vector was successfully constructed. CTL cells were transfected with the established CRSIPR/Cas9 system, and the highest transfection efficiency was up to (28.80±0.62)%. After transfection, the deletion efficiency of CTLA-4 was detected by Flow cytometry. The CTLA-4 expression of CTLA-4 KO CTL group was significantly lower than that of CTL group [(0.91±0.25)% vs (42.70±2.72)%, P<0.05]. In prophylactic assay, the formation rate of colon cancer xenografts in the experimental group was significantly lower than that in the control group (33.33%vs 100%, P<0.05). In treatment assay, the tumor volume in the experimental group was significantly inhibited compared with the control group ([503±23.9] vs [911.2±51.4] mm3, P<0.05), and the survival time of the experimental group was significantly prolonged (median survival time: 78 d vs 42 d, P<0.05); Moreover, the secretion levels of serumTNF-α ([268.93±17.04] pg/ml vs [148.26±20.07] pg/ml, P<0.05) and IFN-γ (315.38±18.67 pg/ml vs 202.92±29.32 pg/ml,P<0.05) in the experimental group were significantly higher than those in the control group. Conclusions: The lentiviral vector CRSIPR/Cas9 system is an effective gene editing method; its successful deletion of CTLA-4 in CTL cells can significantly inhibit the tumor formation rate of colon cancer xenografts in nude mice and enhance the anti-tumor effect of CTL on colon cancer xenografts.
2020, 27(3):228-234.
DOI: 10.3872/j.issn.1007-385X.2020.03.003
Abstract:
Objective: To investigate the effect of Baohuoside-Ⅰ on the proliferation, invasion, migration and apoptosis of colon cancer cell lines SW480 and RKO and the relative mechanism. Methods: Colon cancer cell lines SW480 and RKO were respectively treated with different concentrations of Baohuoside-Ⅰ (0, 5, 10, 20 μg/ml). Cell proliferation was detected by MTT assay; The ability of cell clone formation was tested by cell clone formation experiments; The migration and invasion of cells were detected by Transwell assay;The apoptosis and cell cycle was detected by Flow cytometry; and the protein expression levels of cleaved PARP, cleaved Caspase-3 and Bcl-2 were detected by Western blotting. The effects of Baohuoside-Ⅰ on transcriptome and possible signaling pathways were detected by RNA-Seq technology. Results: Baohuoside-Ⅰ could inhibit the proliferation, invasion and migration of SW480 and RKO cells, and induce cell apoptosis and G0/G1 phase block. Baohuoside-Ⅰ could also up-regulate the protein expressions of cleaved PARP and cleaved Caspase-3 but down-regulate the protein expression of Bcl-2 in SW480 and RKO cell lines. In addition, RNA-Seq data analysis showed that DNA replication and transcription of ERBB signaling pathway related genes were both affected by Baohuoside-Ⅰ. Conclusion: Baohuoside-Ⅰ could induce apoptosis and G0/G1 phase block of colon cancer cell lines SW480 and RKO by affecting the expression of apoptosis related proteins, as well as cellular DNA replication and ERBB signaling pathways, thus inhibiting the malignant phenotypes of SW480 and RKO.
Objective: To investigate the effect of Baohuoside-Ⅰ on the proliferation, invasion, migration and apoptosis of colon cancer cell lines SW480 and RKO and the relative mechanism. Methods: Colon cancer cell lines SW480 and RKO were respectively treated with different concentrations of Baohuoside-Ⅰ (0, 5, 10, 20 μg/ml). Cell proliferation was detected by MTT assay; The ability of cell clone formation was tested by cell clone formation experiments; The migration and invasion of cells were detected by Transwell assay;The apoptosis and cell cycle was detected by Flow cytometry; and the protein expression levels of cleaved PARP, cleaved Caspase-3 and Bcl-2 were detected by Western blotting. The effects of Baohuoside-Ⅰ on transcriptome and possible signaling pathways were detected by RNA-Seq technology. Results: Baohuoside-Ⅰ could inhibit the proliferation, invasion and migration of SW480 and RKO cells, and induce cell apoptosis and G0/G1 phase block. Baohuoside-Ⅰ could also up-regulate the protein expressions of cleaved PARP and cleaved Caspase-3 but down-regulate the protein expression of Bcl-2 in SW480 and RKO cell lines. In addition, RNA-Seq data analysis showed that DNA replication and transcription of ERBB signaling pathway related genes were both affected by Baohuoside-Ⅰ. Conclusion: Baohuoside-Ⅰ could induce apoptosis and G0/G1 phase block of colon cancer cell lines SW480 and RKO by affecting the expression of apoptosis related proteins, as well as cellular DNA replication and ERBB signaling pathways, thus inhibiting the malignant phenotypes of SW480 and RKO.
2020, 27(3):235-241.
DOI: 10.3872/j.issn.1007-385X.2020.03.004
Abstract:
Objective: To design and prepare a novel bi-specific chimeric antigen receptor (CAR) -T cell targeting both CD20 and CD19 antigen on B lymphocyte surface, and to detect its killing effect on B lymphocyte tumors as well as its treatment efficacy on immunodeficiency B-NSG mouse with leukemia. Methods: Bi-specific CAR molecule of CD20 (human originated)/CD19 (murine originated) scFv was constructed and packaged into lentiviral vector in 293 cells, and then transfected into T lymphocytes from healthy donors to prepare BiCAR-T cells. K562-CD19-GFP cells (with positive CD19 expression), K562-CD20-GFP cells (with positive CD20 expression) and Nalm6-Luc-GFP cells expressing luciferase were constructed as target cells. After being co-incubated with above mentioned targets cells, the cytotoxic effects of BiCAR-T cells on target cells were evaluated via LDH release assay, and the secretion of IFN-γ by BiCAR-T cells was evaluated by ELISA. Nalm6-Luc-GFP cells were used to construct the mouse model of leukemia and BiCAR-T cells were transfused via tail veins; the treatment efficacy of BiCAR-T cells on tumor bearing mice was evaluated with small animal imaging method. Results: After 7 days’incubation, the BiCAR-T cells originated from healthy donors amplified about 20-50 times with a positive rate of 10%~92%, indicating successful preparation of BiCAR-T cells. Under an effector∶target ratio of 10∶1, the killing rates of BiCAR-T cells against Nalm-6, K562-CD19-GFP and K562-CD20-GFP cells were significantly higher than that of control cells [(76.7±7.4)% vs (8.7±2.4)%, (93.3±5.2)% vs (46.7±6.2)%, (51.0±0.8) vs (30.7±0.5)%, all P<0.01]. Compared with control group, BiCAR-T cells co-incubated with Nalm-6 cells also secreted significantly more IFN- γ [(872.7±7.7) vs (101.0±5.3)pg/ml, P<0.01). Animal experiment showed that BiCAR-T cells had significant efficacy on B-NSG mice with leukemia; NSG leukemia mice treated with BiCAR-T cells all lived up to 70 days (till they were mercy killed) and leukemia cells disappeared at about 50 days,while the mice in PBS and T lymphocytes group all died at (19±3) d and (20±1) d, respectively. Conclusion: Bi-specific CAR molecules expressing CD19 and CD20 were successfully designed and BiCAR-T cells were successfully prepared. The BiCAR-T cells can effectively kill CD19 and/or CD20 tumor cells and secret large amounts of IFN-γ after co-incubation with target cells, exerting significant treatment efficacy on B-NSG immunodeficiency mouse with leukemia.
Objective: To design and prepare a novel bi-specific chimeric antigen receptor (CAR) -T cell targeting both CD20 and CD19 antigen on B lymphocyte surface, and to detect its killing effect on B lymphocyte tumors as well as its treatment efficacy on immunodeficiency B-NSG mouse with leukemia. Methods: Bi-specific CAR molecule of CD20 (human originated)/CD19 (murine originated) scFv was constructed and packaged into lentiviral vector in 293 cells, and then transfected into T lymphocytes from healthy donors to prepare BiCAR-T cells. K562-CD19-GFP cells (with positive CD19 expression), K562-CD20-GFP cells (with positive CD20 expression) and Nalm6-Luc-GFP cells expressing luciferase were constructed as target cells. After being co-incubated with above mentioned targets cells, the cytotoxic effects of BiCAR-T cells on target cells were evaluated via LDH release assay, and the secretion of IFN-γ by BiCAR-T cells was evaluated by ELISA. Nalm6-Luc-GFP cells were used to construct the mouse model of leukemia and BiCAR-T cells were transfused via tail veins; the treatment efficacy of BiCAR-T cells on tumor bearing mice was evaluated with small animal imaging method. Results: After 7 days’incubation, the BiCAR-T cells originated from healthy donors amplified about 20-50 times with a positive rate of 10%~92%, indicating successful preparation of BiCAR-T cells. Under an effector∶target ratio of 10∶1, the killing rates of BiCAR-T cells against Nalm-6, K562-CD19-GFP and K562-CD20-GFP cells were significantly higher than that of control cells [(76.7±7.4)% vs (8.7±2.4)%, (93.3±5.2)% vs (46.7±6.2)%, (51.0±0.8) vs (30.7±0.5)%, all P<0.01]. Compared with control group, BiCAR-T cells co-incubated with Nalm-6 cells also secreted significantly more IFN- γ [(872.7±7.7) vs (101.0±5.3)pg/ml, P<0.01). Animal experiment showed that BiCAR-T cells had significant efficacy on B-NSG mice with leukemia; NSG leukemia mice treated with BiCAR-T cells all lived up to 70 days (till they were mercy killed) and leukemia cells disappeared at about 50 days,while the mice in PBS and T lymphocytes group all died at (19±3) d and (20±1) d, respectively. Conclusion: Bi-specific CAR molecules expressing CD19 and CD20 were successfully designed and BiCAR-T cells were successfully prepared. The BiCAR-T cells can effectively kill CD19 and/or CD20 tumor cells and secret large amounts of IFN-γ after co-incubation with target cells, exerting significant treatment efficacy on B-NSG immunodeficiency mouse with leukemia.
2020, 27(3):242-247.
DOI: 10.3872/j.issn.1007-385X.2020.03.005
Abstract:
Objective: To investigate the effect of salivary adenoid cystic carcinoma (SACC) derived exosomes on PD-L1 expression in fibroblasts. Methods: Exosomes of SACC-83 cell line were extracted by exosome isolation kit, and its particle size, density and phenotypes were identified by electron microscope. After being labeled with PKH67 fluorescence, exosomes were co-incubated with HPLF to observe whether the exosomes could be ingested by fibroblasts under confocal microscope. After co-incubation with the exosomes,the differentially expressed genes (DEGs) in HPLF cells were detected by whole transcriptome sequencing. GO analysis together with KEGG enrichment analysis was used to clarify the biological functions and related signaling pathways related with the DEGs. qPCR,Western blotting and Flow cytometry were used to detect the effect of tumor exosomes on the mRNA and protein expressions of PD-L1,LAG3 and IDO1 in HPLF fibroblasts. Results: SACC exosomes specifically expressed CD63, CD81 and TSG101 molecules, and could be ingested by fibroblasts. After the treatment of fibroblasts by exosomes, the expression of PD-L1 molecule was significantly up-regulated (Fold change=10.19), and the DEGs were significantly enriched in the immune response signaling pathways, such as TNF, NF-κB and cGAS-String pathway, etc. In vitro experiments showed that exosomes could significantly promote the expression of PD-L1 in HPLF cells at both mRNA and protein levels (24.7±4.75 vs 1.03±0.11,P<0.05). Conclusion: Exosomes of SACC can promote the expression of immunocheckpoint ligand PD-L1 in fibroblasts.
Objective: To investigate the effect of salivary adenoid cystic carcinoma (SACC) derived exosomes on PD-L1 expression in fibroblasts. Methods: Exosomes of SACC-83 cell line were extracted by exosome isolation kit, and its particle size, density and phenotypes were identified by electron microscope. After being labeled with PKH67 fluorescence, exosomes were co-incubated with HPLF to observe whether the exosomes could be ingested by fibroblasts under confocal microscope. After co-incubation with the exosomes,the differentially expressed genes (DEGs) in HPLF cells were detected by whole transcriptome sequencing. GO analysis together with KEGG enrichment analysis was used to clarify the biological functions and related signaling pathways related with the DEGs. qPCR,Western blotting and Flow cytometry were used to detect the effect of tumor exosomes on the mRNA and protein expressions of PD-L1,LAG3 and IDO1 in HPLF fibroblasts. Results: SACC exosomes specifically expressed CD63, CD81 and TSG101 molecules, and could be ingested by fibroblasts. After the treatment of fibroblasts by exosomes, the expression of PD-L1 molecule was significantly up-regulated (Fold change=10.19), and the DEGs were significantly enriched in the immune response signaling pathways, such as TNF, NF-κB and cGAS-String pathway, etc. In vitro experiments showed that exosomes could significantly promote the expression of PD-L1 in HPLF cells at both mRNA and protein levels (24.7±4.75 vs 1.03±0.11,P<0.05). Conclusion: Exosomes of SACC can promote the expression of immunocheckpoint ligand PD-L1 in fibroblasts.
2020, 27(3):248-254.
DOI: 10.3872/j.issn.1007-385X.2020.03.006
Abstract:
Objective：To explore the targeting relationship between miR-377-5p and hypoxia inducible factor-1 (HIF-1α), and investigate the regulatory effect of miR-377-5p on proliferation, invasion and epithelial-mesenchymal transition (EMT) of hepatocellular carcinoma (HCC) cells through vascular endothelial growth factor (VEGF) signaling pathway. Methods：The expression of miR-377-5p in 35 pairs of human HCC tissues and para-cancerous tissues was detected by qPCR. Then, HepG2 cells were divided into control group, mimic-NC group and miR-377-5p mimic group. qPCR was used to detect the transfection efficiency; the effects of miR-377-5p over-expression on proliferation and invasion of HepG2 cells were examined by EdU staining and Transwell assay, respectively; and the effect of miR-377-5p over-expression on the expressions of proliferation-related protein Ki-67, proliferating cell nuclear antigen (PCNA) and epithelial-mesenchymal transition (EMT) markers (E-cadherin and N-cadherin) were detected by Western blotting (WB);the effect of miR-377-5p over-expression on the expression of hypoxia inducible factor-1α (HIF-1α ) in HepG2 cells was detected by qPCR and WB; and the targeting relationship between miR-377-5p and HIF-1α gene was determined by Luciferase reporter gene assay.Results: The expression of miR-377-5p in HCC tissues was significantly lower than that in para-cancerous tissues (P<0.01). Compared with the control group, the expression of miR-377-5p in HepG2 cells of miR-377-5p mimic group elevated significantly, and the proliferation,invasion and the expression of N-caderin proteins decreased, significantly (all P<0.01), while the expression of E-caderin increased significantly (P<0.01). At the same time, the mRNA and protein expressions of HIF-1α in miR-377-5p mimic group decreased significantly (P<0.01 or P<0.05). miR-377-5p targetedly inhibited the expression of HIF-1α gene and suppressed the activation of VEGF pathway (all P<0.05). Conclusion: miR-377-5p inhibits the proliferation, invasion and EMT of HepG2 cells via targetedly inhibiting HIF-1α expression and suppressing the activation of VEGF signaling pathway.
Objective：To explore the targeting relationship between miR-377-5p and hypoxia inducible factor-1 (HIF-1α), and investigate the regulatory effect of miR-377-5p on proliferation, invasion and epithelial-mesenchymal transition (EMT) of hepatocellular carcinoma (HCC) cells through vascular endothelial growth factor (VEGF) signaling pathway. Methods：The expression of miR-377-5p in 35 pairs of human HCC tissues and para-cancerous tissues was detected by qPCR. Then, HepG2 cells were divided into control group, mimic-NC group and miR-377-5p mimic group. qPCR was used to detect the transfection efficiency; the effects of miR-377-5p over-expression on proliferation and invasion of HepG2 cells were examined by EdU staining and Transwell assay, respectively; and the effect of miR-377-5p over-expression on the expressions of proliferation-related protein Ki-67, proliferating cell nuclear antigen (PCNA) and epithelial-mesenchymal transition (EMT) markers (E-cadherin and N-cadherin) were detected by Western blotting (WB);the effect of miR-377-5p over-expression on the expression of hypoxia inducible factor-1α (HIF-1α ) in HepG2 cells was detected by qPCR and WB; and the targeting relationship between miR-377-5p and HIF-1α gene was determined by Luciferase reporter gene assay.Results: The expression of miR-377-5p in HCC tissues was significantly lower than that in para-cancerous tissues (P<0.01). Compared with the control group, the expression of miR-377-5p in HepG2 cells of miR-377-5p mimic group elevated significantly, and the proliferation,invasion and the expression of N-caderin proteins decreased, significantly (all P<0.01), while the expression of E-caderin increased significantly (P<0.01). At the same time, the mRNA and protein expressions of HIF-1α in miR-377-5p mimic group decreased significantly (P<0.01 or P<0.05). miR-377-5p targetedly inhibited the expression of HIF-1α gene and suppressed the activation of VEGF pathway (all P<0.05). Conclusion: miR-377-5p inhibits the proliferation, invasion and EMT of HepG2 cells via targetedly inhibiting HIF-1α expression and suppressing the activation of VEGF signaling pathway.
2020, 27(3):255-260.
DOI: 10.3872/j.issn.1007-385X.2020.03.007
Abstract:
Objective: To explore the effect of interfering insulin-like growth factors-1 receptors (IGF-1R) by small interfering RNA (siRNA) on cell cycle and apoptosis of hypoxic hepatocellular carcinoma HepG2 cells. Methods: The hypoxic hepatocellular carcinoma model was established via cobalt chloride treatment. Three siRNAs targeting IGF1R gene and one negative control siRNA were designed and synthesized. They were transfected into hypoxic HepG2 cells, and 24 h later, the transfection efficiency was detected by fluorescent microscopy. The protein expression of IFG-1R was detected with Western blotting (WB) to screen the siRNA with highest transfection efficacy. The selected siRNA was used to transfect hypoxic HepG2 cells. The proliferation of hypoxic HepG2 cells was determined by MTT assay. Cell cycle distribution and apoptosis were analyzed by Flow cytometry. WB was performed to detect the protein expressions of CDK1, CDK2 and Caspase-3 in HepG2 cells. Results: The hypoxic hepatocellular carcinoma model was successfully established. IGF-1R-siRNA-2 showed the most effective interference efficiency and the most significant knockdown of IGF-1R (all P<0.01). The proliferation of HepG2 cells transfected with IGF-1R siRNA-2 was significantly suppressed (P<0.05 or P<0.01), the cell cycle was blocked at G0/G1 phase (P<0.05), and the apoptosis rate was increased up to (25.3±1.3)% P<0.01). In the meanwhile,the expressions of CDK1 and CDK2 were decreased and the expression of Caspase-3 was increased in hypoxic HepG2 cells after IGF-1R knockdown (P<0.05). Conclusion: Interfering IGF-1R by siRNA inhibits the malignant biological behaviors of hypoxic HepG2 cells via regulating cell cycle and apoptosis-related proteins. IGF-1R may be a potential target for the treatment of HCC.
Objective: To explore the effect of interfering insulin-like growth factors-1 receptors (IGF-1R) by small interfering RNA (siRNA) on cell cycle and apoptosis of hypoxic hepatocellular carcinoma HepG2 cells. Methods: The hypoxic hepatocellular carcinoma model was established via cobalt chloride treatment. Three siRNAs targeting IGF1R gene and one negative control siRNA were designed and synthesized. They were transfected into hypoxic HepG2 cells, and 24 h later, the transfection efficiency was detected by fluorescent microscopy. The protein expression of IFG-1R was detected with Western blotting (WB) to screen the siRNA with highest transfection efficacy. The selected siRNA was used to transfect hypoxic HepG2 cells. The proliferation of hypoxic HepG2 cells was determined by MTT assay. Cell cycle distribution and apoptosis were analyzed by Flow cytometry. WB was performed to detect the protein expressions of CDK1, CDK2 and Caspase-3 in HepG2 cells. Results: The hypoxic hepatocellular carcinoma model was successfully established. IGF-1R-siRNA-2 showed the most effective interference efficiency and the most significant knockdown of IGF-1R (all P<0.01). The proliferation of HepG2 cells transfected with IGF-1R siRNA-2 was significantly suppressed (P<0.05 or P<0.01), the cell cycle was blocked at G0/G1 phase (P<0.05), and the apoptosis rate was increased up to (25.3±1.3)% P<0.01). In the meanwhile,the expressions of CDK1 and CDK2 were decreased and the expression of Caspase-3 was increased in hypoxic HepG2 cells after IGF-1R knockdown (P<0.05). Conclusion: Interfering IGF-1R by siRNA inhibits the malignant biological behaviors of hypoxic HepG2 cells via regulating cell cycle and apoptosis-related proteins. IGF-1R may be a potential target for the treatment of HCC.
8 Effect and molecular mechanism of Lin28 on 5-Fu sensitivity of hepatocellular carcinoma HepG2 cells
2020, 27(3):261-266.
DOI: 10.3872/j.issn.1007-385X.2020.03.008
Abstract:
Objective: To investigate the effect and mechanism of RNA binding protein Lin28 on the 5-fluorouracil (5-Fu) sensitivity of HepG2 cells. Methods: HepG2 cells were transfected with plasmid pcDNA3.1-Lin28 or si-Lin28 (small interfering RNA of Lin28).qPCR and Western blotting were used to detect the expression of Lin28 in HepG2 cells after transfection. Changes of cell proliferation in transfected cells after 5-Fu treatment was detected by CCK8 assay and the 50% inhibitory concentration (IC50) was calculated. Flow cytometry was used to detect apoptotic rate after 5-Fu treatment and the expression of apoptosis-related protein was assayed by Western blotting. The mRNA expressions of drug-resistant miRNAs (let-7a and let-7b), as well as cancer stem cell markers (Oct4, Nanog and Sox2) after transfection were detected by qPCR. Results: As compared to the HepG2/Vector cells, the mRNA and protein expressions of Lin28 were significantly up-regulated in HepG2/Lin28 cells (P<0.05 or P<0.01). Over-expression of Lin28 significantly suppressed the sensitivity of HepG2 cells to 5-Fu (IC50 elevated obviously, P<0.05) and significantly increased cell proliferation while decreased apoptotic rate and expression of apoptotic-related protein caspase-3 (all P<0.01). As compared to si-control group, expression of Lin28 in HepG2/si-Lin28 cells was significantly down-regulated (P<0.01). Lin28 knockdown significantly reduced cell proliferation and IC50 of 5-Fu (all P<0.01) but increased apoptotic rate and expression of apoptosis-related protein (P<0.01). Compared with HepG2/Vector group, expressions of let-7a and let-7b, as well as cancer stem cell markers (Oct4, Nanog and Sox2) were significantly increased in HepG2/Lin28 cells (all P<0.01); while these molecules were significantly decreased in HepG2/si-Lin28 cells as comparing to si-control group (all P<0.01). Conclusion: Lin28 can modulate the chemosensitivity of HepG2 cells by regulating the expression of miRNAs and the formation of cancer stem cells. Targeting Lin28 might be a promising approach to improve the chemotherapy efficacy in HCC.
Objective: To investigate the effect and mechanism of RNA binding protein Lin28 on the 5-fluorouracil (5-Fu) sensitivity of HepG2 cells. Methods: HepG2 cells were transfected with plasmid pcDNA3.1-Lin28 or si-Lin28 (small interfering RNA of Lin28).qPCR and Western blotting were used to detect the expression of Lin28 in HepG2 cells after transfection. Changes of cell proliferation in transfected cells after 5-Fu treatment was detected by CCK8 assay and the 50% inhibitory concentration (IC50) was calculated. Flow cytometry was used to detect apoptotic rate after 5-Fu treatment and the expression of apoptosis-related protein was assayed by Western blotting. The mRNA expressions of drug-resistant miRNAs (let-7a and let-7b), as well as cancer stem cell markers (Oct4, Nanog and Sox2) after transfection were detected by qPCR. Results: As compared to the HepG2/Vector cells, the mRNA and protein expressions of Lin28 were significantly up-regulated in HepG2/Lin28 cells (P<0.05 or P<0.01). Over-expression of Lin28 significantly suppressed the sensitivity of HepG2 cells to 5-Fu (IC50 elevated obviously, P<0.05) and significantly increased cell proliferation while decreased apoptotic rate and expression of apoptotic-related protein caspase-3 (all P<0.01). As compared to si-control group, expression of Lin28 in HepG2/si-Lin28 cells was significantly down-regulated (P<0.01). Lin28 knockdown significantly reduced cell proliferation and IC50 of 5-Fu (all P<0.01) but increased apoptotic rate and expression of apoptosis-related protein (P<0.01). Compared with HepG2/Vector group, expressions of let-7a and let-7b, as well as cancer stem cell markers (Oct4, Nanog and Sox2) were significantly increased in HepG2/Lin28 cells (all P<0.01); while these molecules were significantly decreased in HepG2/si-Lin28 cells as comparing to si-control group (all P<0.01). Conclusion: Lin28 can modulate the chemosensitivity of HepG2 cells by regulating the expression of miRNAs and the formation of cancer stem cells. Targeting Lin28 might be a promising approach to improve the chemotherapy efficacy in HCC.
9 circ_0005075 promotes proliferation and invasion of liver cancer HCCC9810 cells by sponging miR-335
2020, 27(3):267-272.
DOI: 10.3872/j.issn.1007-385X.2020.03.009
Abstract:
Objective: To explore the effect of circ_0005075 on the proliferation and invasion of liver cancer cells and its underlying mechanism. Methods: A total of 35 cases of cancer tissues and corresponding para-cancerous tissues from liver cancer patients, who underwent surgical resection in Tangshan Workers’Hospital from March 2015 to March 2018, were collected for this study. qPCR was used to detect the expression levels of circ_0005075 and miR-335 in liver cancer tissues, para-cancerous tissues, liver cancer cell lines (HCCC9810, HepG2, HLE and hepatic epithelial THLE-3 cells). Dual luciferase reporter gene assay was used to verify the targeting relationship among circ 0005075, mir-335 and CCND1. By using liposome-mediated method, Sh-circ_0005075, miR-335 mimics, miR-335 mimics+pcDNA-CCND1, sh-circ_0005075+pcDNA-CCND1, pcDNA-circ_0005075+miR-335 mimics, sh-CCND1+pcDNA-circ_0005075 were transfected into HCCC9810 cells, respectively. The effects of circ_0005075/miR-335/CCND1 molecular axis on the proliferation and invasion of HCCC9810 cells were detected by MTT and Transwell methods. Results: circ_0005075 was highly expressed in liver cancer tissues and cell lines (P<0.01) ,and the highest expression in HCCC9810 cells (P<0.05). Dual luciferase reporter gene results showed that circ_0005075 negatively regulated miR-335 (P<0.05), and CCND1 was a target gene of miR-335 (P<0.05). Further experiments proved that knockdown of circ_0005075 or overexpression of miR-335 could inhibit the proliferation and invasion of HCCC9810 cells by regulating CCND1（P<0.05 or P<0.01）. Conclusion: Circ_0005075 upregulates the expression level of CCND1 by sponging miR-335, thereby promoting the proliferation and invasion of HCCC9810 cells.
Objective: To explore the effect of circ_0005075 on the proliferation and invasion of liver cancer cells and its underlying mechanism. Methods: A total of 35 cases of cancer tissues and corresponding para-cancerous tissues from liver cancer patients, who underwent surgical resection in Tangshan Workers’Hospital from March 2015 to March 2018, were collected for this study. qPCR was used to detect the expression levels of circ_0005075 and miR-335 in liver cancer tissues, para-cancerous tissues, liver cancer cell lines (HCCC9810, HepG2, HLE and hepatic epithelial THLE-3 cells). Dual luciferase reporter gene assay was used to verify the targeting relationship among circ 0005075, mir-335 and CCND1. By using liposome-mediated method, Sh-circ_0005075, miR-335 mimics, miR-335 mimics+pcDNA-CCND1, sh-circ_0005075+pcDNA-CCND1, pcDNA-circ_0005075+miR-335 mimics, sh-CCND1+pcDNA-circ_0005075 were transfected into HCCC9810 cells, respectively. The effects of circ_0005075/miR-335/CCND1 molecular axis on the proliferation and invasion of HCCC9810 cells were detected by MTT and Transwell methods. Results: circ_0005075 was highly expressed in liver cancer tissues and cell lines (P<0.01) ,and the highest expression in HCCC9810 cells (P<0.05). Dual luciferase reporter gene results showed that circ_0005075 negatively regulated miR-335 (P<0.05), and CCND1 was a target gene of miR-335 (P<0.05). Further experiments proved that knockdown of circ_0005075 or overexpression of miR-335 could inhibit the proliferation and invasion of HCCC9810 cells by regulating CCND1（P<0.05 or P<0.01）. Conclusion: Circ_0005075 upregulates the expression level of CCND1 by sponging miR-335, thereby promoting the proliferation and invasion of HCCC9810 cells.
2020, 27(3):273-281.
DOI: 10.3872/j.issn.1007-385X.2020.03.010
Abstract:
Objective: To investigate the effect of long non-coding RNA (lncRNA) lung cancer associated transcript 1 (LUCAT1) on proliferation and migration of clear cell renal cell carcinoma (ccRCC) 786-O cells and the underlying mechanism. Methods: A total of 40 pairs of pathologically confirmed tumor tissues and corresponding adjacent normal tissues from ccRCC patients, who underwent surgical resection in the Department of Urology, the First People's Hospital of Yichang during June 2013 and June 2017, were selected for this study. ccRCC cell lines (786-O, ACHN, UM-RC-2) and normal renal epithelial KiMA cells were also used in this study. qPCR was used to detect the mRNA expressions of LUCAT1, miR-199a-5p and hypoxia inducible fator 1α (HIF-1α) in above mentioned tissues and cell lines; CCK-8 assay was used to evaluate the proliferation of 786-O cells; Transwell assay was used to evaluate the migration of 786-O cells; Dual luciferase reporter gene assay was performed to validate the relationship between LUCAT1 and miR-199a-5p; and Western blotting was conducted to detect the effect of LUCAT1 and miR-199a-5p on the protein expression of HIF-1α . Results:LUCAT1 was significantly up-regulated in ccRCC tissues and cell lines (all P<0.01), and its knockdown significantly inhibited the proliferation and migration of 786-O cells (all P<0.01). miR-199a-5p was low-expressed in ccRCC tissues and cell lines (all P<0.01), Star‐Base analysis showed that LUCAT1 contained a conserved target site for miR-199a-5p. miR-199a-5p exerted significant suppression on the luciferase activity of LUCAT1-Wt (P<0.01), and LUCAT1 knockdown significantly reduced miR-199a-5p expression (P<0.01).LUCAT1 was low-expressed in 786-O cells transfected with miR-199a-5p mimics, however, it was attenuated after co-transfection with LUCAT1. The mRNA and protein expressions of HIF-1α in 786-O cells transfected with miR-199a-5p mimics were up-regulated,which was then reversed by LUCAT1 over-expression (P<0.05 or P<0.01). miR-199a-5p over-expression suppressed the proliferation and migration of 786-O cells, which was partially attenuated by LUCAT1 transfection (P<0.05 or P<0.01). Conclusion: LUCAT1 exerts oncogenic function in ccRCC via regulating miR-199a-5p/HIF-1α axis.
Objective: To investigate the effect of long non-coding RNA (lncRNA) lung cancer associated transcript 1 (LUCAT1) on proliferation and migration of clear cell renal cell carcinoma (ccRCC) 786-O cells and the underlying mechanism. Methods: A total of 40 pairs of pathologically confirmed tumor tissues and corresponding adjacent normal tissues from ccRCC patients, who underwent surgical resection in the Department of Urology, the First People's Hospital of Yichang during June 2013 and June 2017, were selected for this study. ccRCC cell lines (786-O, ACHN, UM-RC-2) and normal renal epithelial KiMA cells were also used in this study. qPCR was used to detect the mRNA expressions of LUCAT1, miR-199a-5p and hypoxia inducible fator 1α (HIF-1α) in above mentioned tissues and cell lines; CCK-8 assay was used to evaluate the proliferation of 786-O cells; Transwell assay was used to evaluate the migration of 786-O cells; Dual luciferase reporter gene assay was performed to validate the relationship between LUCAT1 and miR-199a-5p; and Western blotting was conducted to detect the effect of LUCAT1 and miR-199a-5p on the protein expression of HIF-1α . Results:LUCAT1 was significantly up-regulated in ccRCC tissues and cell lines (all P<0.01), and its knockdown significantly inhibited the proliferation and migration of 786-O cells (all P<0.01). miR-199a-5p was low-expressed in ccRCC tissues and cell lines (all P<0.01), Star‐Base analysis showed that LUCAT1 contained a conserved target site for miR-199a-5p. miR-199a-5p exerted significant suppression on the luciferase activity of LUCAT1-Wt (P<0.01), and LUCAT1 knockdown significantly reduced miR-199a-5p expression (P<0.01).LUCAT1 was low-expressed in 786-O cells transfected with miR-199a-5p mimics, however, it was attenuated after co-transfection with LUCAT1. The mRNA and protein expressions of HIF-1α in 786-O cells transfected with miR-199a-5p mimics were up-regulated,which was then reversed by LUCAT1 over-expression (P<0.05 or P<0.01). miR-199a-5p over-expression suppressed the proliferation and migration of 786-O cells, which was partially attenuated by LUCAT1 transfection (P<0.05 or P<0.01). Conclusion: LUCAT1 exerts oncogenic function in ccRCC via regulating miR-199a-5p/HIF-1α axis.
2020, 27(3):282-288.
DOI: 10.3872/j.issn.1007-385X.2020.03.011
Abstract:
Objective: To explore the roles and mechanisms of long non-coding RNA (lncRNA) small nucleolar RNA host gene 6 (SNHG6) in promoting invasion and metastasis of esophageal squamous carcinoma (ESCC). Methods: Real time quantitative polymerase chain reaction (qPCR) was used to detect the expression of SNHG6 in ESCC and matched para-carcinoma tissues. Reverse transcription-polymerase chain reaction (RT-PCR) was performed to detect the expression of SNHG6 in ESCC cell lines (TE1, Yes-2,Eca9706 and Kyse150). Then, TE1 cell line which harbored highest expression of SNHG6 was used in following experiments. siRNAs were used to knock down the expression of SNHG6. Clone formation, wound-healing and transwell assay were used to detect the abilities of proliferation, migration and invasion of TE1 cells, respectively.Western blotting was used to detect the expressions of MMP-2,MMP-9 andZEB1 protein before and after knockdown of SNHG6 in TE1 cells. Results: SNHG6 was highly expressed in ESCC tissues,compared to para-carcinoma tissues (P<0.01). The expression of SNHG6 was significantly decreased after transfection of SNHG6-siRNA (all P<0.01). The abilities of proliferation, migration and invasion of TE1 cells in si-SNHG6-1 and si-SNHG6-2 group were significantly lower than those in the control group (all P<0.01). The expressions of ZEB1, MMP-2 and MMP-9 in si-SNHG6-1 and si-SNHG6-2 group were significantly lower than those in the control group (all P<0.05). Conclusion: SNHG6 is highly expressed in ESCC tissues and promotes the malignant biological behavior of ESCC cells. Its mechanism of promoting the occurrence and development of ESCC may be related to the upregulation of ZEB1 expression.
Objective: To explore the roles and mechanisms of long non-coding RNA (lncRNA) small nucleolar RNA host gene 6 (SNHG6) in promoting invasion and metastasis of esophageal squamous carcinoma (ESCC). Methods: Real time quantitative polymerase chain reaction (qPCR) was used to detect the expression of SNHG6 in ESCC and matched para-carcinoma tissues. Reverse transcription-polymerase chain reaction (RT-PCR) was performed to detect the expression of SNHG6 in ESCC cell lines (TE1, Yes-2,Eca9706 and Kyse150). Then, TE1 cell line which harbored highest expression of SNHG6 was used in following experiments. siRNAs were used to knock down the expression of SNHG6. Clone formation, wound-healing and transwell assay were used to detect the abilities of proliferation, migration and invasion of TE1 cells, respectively.Western blotting was used to detect the expressions of MMP-2,MMP-9 andZEB1 protein before and after knockdown of SNHG6 in TE1 cells. Results: SNHG6 was highly expressed in ESCC tissues,compared to para-carcinoma tissues (P<0.01). The expression of SNHG6 was significantly decreased after transfection of SNHG6-siRNA (all P<0.01). The abilities of proliferation, migration and invasion of TE1 cells in si-SNHG6-1 and si-SNHG6-2 group were significantly lower than those in the control group (all P<0.01). The expressions of ZEB1, MMP-2 and MMP-9 in si-SNHG6-1 and si-SNHG6-2 group were significantly lower than those in the control group (all P<0.05). Conclusion: SNHG6 is highly expressed in ESCC tissues and promotes the malignant biological behavior of ESCC cells. Its mechanism of promoting the occurrence and development of ESCC may be related to the upregulation of ZEB1 expression.
12 CARD10 promotes apoptosis inhibition of hepatocellular carcinoma cells by activating NF-κB pathway
2020, 27(3):289-294.
DOI: 10.3872/j.issn.1007-385X.2020.03.012
Abstract:
Objective: To investigate the expression of CARD10 in hepatocellular carcinoma (HCC) tissues, and the roles of CARD10 in HCC progression especially apoptosis inhibition. Methods: The expression of CARD10 was examined in pared non-tumor liver tissues and HCC tissues using qRT-PCR, and their correlation with HCC TNM stage was analyzed using Spearman’s rank correlation assay in SPSS 17.0. In HCC cells with CARD10 overexpression or knockdown, cytometry using Annexin-V/PI labeling was used to measure apoptosis, and Western blotting was used to determine the activation of NF- κB pathway. Results: CARD10 expression was significantly increased in HCC tissues as compared to that in pared non-tumor liver tissues (P<0.01), and the increased CARD10 in HCC was positively correlated with TNM staging (P<0.01). The apoptosis of HCC cell lines SMMC-7721 and BEL-7402 was inhibited by CARD10 overexpression while promoted by CARD10 knockdown, and the pro-survival NF- κB pathway was also enhanced by CARD 10 over-expression while suppressed by CARD10 knockdown. Conclusion: CARD10 expression is increased in HCC tissues and positively correlated with HCC progression. CARD10 inhibits HCC apoptosis by promoting the activation of NF-κB pathway.
Objective: To investigate the expression of CARD10 in hepatocellular carcinoma (HCC) tissues, and the roles of CARD10 in HCC progression especially apoptosis inhibition. Methods: The expression of CARD10 was examined in pared non-tumor liver tissues and HCC tissues using qRT-PCR, and their correlation with HCC TNM stage was analyzed using Spearman’s rank correlation assay in SPSS 17.0. In HCC cells with CARD10 overexpression or knockdown, cytometry using Annexin-V/PI labeling was used to measure apoptosis, and Western blotting was used to determine the activation of NF- κB pathway. Results: CARD10 expression was significantly increased in HCC tissues as compared to that in pared non-tumor liver tissues (P<0.01), and the increased CARD10 in HCC was positively correlated with TNM staging (P<0.01). The apoptosis of HCC cell lines SMMC-7721 and BEL-7402 was inhibited by CARD10 overexpression while promoted by CARD10 knockdown, and the pro-survival NF- κB pathway was also enhanced by CARD 10 over-expression while suppressed by CARD10 knockdown. Conclusion: CARD10 expression is increased in HCC tissues and positively correlated with HCC progression. CARD10 inhibits HCC apoptosis by promoting the activation of NF-κB pathway.
2020, 27(3):295-301.
DOI: 10.3872/j.issn.1007-385X.2020.03.013
Abstract:
Objective: To investigate the characteristics and clinical significance of the immunomicroenvironment typing based on the expression of programmed death-ligand 1 (PD-L1) and the infiltration of CD8+ T cells in the stroma in patients with non-small cell lung cancer (NSCLC). Methods: Paraffin tissue specimens and relevant clinicopathological data of 74 NSCLC patients admitted to our hospital from January 2016 to July 2018 were collected. All patients received EGFR gene test, and none received radiotherapy, chemotherapy or targeted therapy. Immunohistochemistry was used to detect the expression of PD-L1 in tissues and the infiltration of CD8+T cells in interstitium, and the relationship between PD-L1, CD8+T cells, and the immune microenvironment typing based on both, and the pathological parameters and the survival of patients was analyzed. Results: PD-L1 expression in the primary tumor of NSCLC patients showed statistical differences in gender, pathological type, smoking history, EFGR gene mutation status (P<0.05). The infiltration of CD8+ T lymphocytes in tumor microenvironment showed statistically significant differences in different TNM stage and lymph node metastasis (P<0.05), PD-L1 expression was significantly correlated with EGFR mutation (P=0.000), while CD8+T lymphocyte infiltration was not correlated with EGFR mutation (P=0.605). The immunomicroenvironment of EGFR wild-type patients was mainly (CD8+PD-L1+) (type I), and the mutants were mainly (CD8-PD-L1-) (type II) and (CD8+PD-L1-) (type IV). The distribution of immune microenvironmental typing in each group with different EGFR mutation, smoking history and pathological differentiation degree was significantly different (P<0.05) and significantly correlated with EGFR mutation (P<0.05). Follow-up showed that the patients with disease-free survival, recurrence and metastasis and death were the most in type I, type II and type I, respectively. Conclusions: In this study,the distribution of tumor immunomicroenvironmental typing in NSCLC patients was mainly the highest in type I and the lowest in type Ⅲ, which was related to EGFR mutation, smoking history and pathological differentiation. Patients with EGFR mutations were mainly of type Ⅱand type Ⅳ, and were associated with low expression of PD-L1.
Objective: To investigate the characteristics and clinical significance of the immunomicroenvironment typing based on the expression of programmed death-ligand 1 (PD-L1) and the infiltration of CD8+ T cells in the stroma in patients with non-small cell lung cancer (NSCLC). Methods: Paraffin tissue specimens and relevant clinicopathological data of 74 NSCLC patients admitted to our hospital from January 2016 to July 2018 were collected. All patients received EGFR gene test, and none received radiotherapy, chemotherapy or targeted therapy. Immunohistochemistry was used to detect the expression of PD-L1 in tissues and the infiltration of CD8+T cells in interstitium, and the relationship between PD-L1, CD8+T cells, and the immune microenvironment typing based on both, and the pathological parameters and the survival of patients was analyzed. Results: PD-L1 expression in the primary tumor of NSCLC patients showed statistical differences in gender, pathological type, smoking history, EFGR gene mutation status (P<0.05). The infiltration of CD8+ T lymphocytes in tumor microenvironment showed statistically significant differences in different TNM stage and lymph node metastasis (P<0.05), PD-L1 expression was significantly correlated with EGFR mutation (P=0.000), while CD8+T lymphocyte infiltration was not correlated with EGFR mutation (P=0.605). The immunomicroenvironment of EGFR wild-type patients was mainly (CD8+PD-L1+) (type I), and the mutants were mainly (CD8-PD-L1-) (type II) and (CD8+PD-L1-) (type IV). The distribution of immune microenvironmental typing in each group with different EGFR mutation, smoking history and pathological differentiation degree was significantly different (P<0.05) and significantly correlated with EGFR mutation (P<0.05). Follow-up showed that the patients with disease-free survival, recurrence and metastasis and death were the most in type I, type II and type I, respectively. Conclusions: In this study,the distribution of tumor immunomicroenvironmental typing in NSCLC patients was mainly the highest in type I and the lowest in type Ⅲ, which was related to EGFR mutation, smoking history and pathological differentiation. Patients with EGFR mutations were mainly of type Ⅱand type Ⅳ, and were associated with low expression of PD-L1.
14 Preliminary bioinformatics analyses of the expression and function of AGR2 in human breast cancer
2020, 27(3):302-308.
DOI: 10.3872/j.issn.1007-385X.2020.03.014
Abstract:
Objective: To explore the expression of AGR2 gene in breast cancer as well as to predict its relevant biological functions and molecular signaling pathways with bioinformatics tool. Methods: The expression of AGR2 in breast cancer tissues and normal tissues was analyzed in Oncomine and GEPIA databases, and the expression of AGR2 in breast cancer cell lines was evaluated in CCLE database. Meanwhile, HPA database was used to analyze the expression of AGR2 protein in normal and breast cancer tissues. Besides,the gene expression microarray data download from CCLE database was analyzed by using R software to obtain genes co-expressed with AGR2. Functional annotation of AGR2 co-expressed genes was performed by using GO Enrichment and KEGG pathway analyses.Results: Oncomine and GEPIA databases showed that AGR2 gene was highly expressed in breast cancer tissues, and CCLE database analysis showed that AGR2 was highly expressed in all breast cancer cell lines. Immunohistochemistry results from the HPA database showed that the expression of AGR2 protein was significantly higher in breast cancer tissues compared with normal tissues. A total of 946 genes co-expressed with AGR2 in breast cancer were screened out with the R software. With the GO function Enrichment analysis,the co-expressed genes were demonstrated to be mainly involved in biological functions, such as protein localization to cell periphery,protein localization to plasma membrane, cell junction assembly, cell-substrate adhesion, and cell junction organization etc. In addition,the KEGG analysis results showed that co-expressed genes were mainly involved in the progression of gastric cancer and breast cancer,and were associated with proteoglycans in cancer, as well as proline, leucine and isoleucine degradation pathways. Conclusions: AGR2 is highly expressed in breast cancer tissues, which may be a potential oncogenic gene and a new therapeutic target of breast cancer.
Objective: To explore the expression of AGR2 gene in breast cancer as well as to predict its relevant biological functions and molecular signaling pathways with bioinformatics tool. Methods: The expression of AGR2 in breast cancer tissues and normal tissues was analyzed in Oncomine and GEPIA databases, and the expression of AGR2 in breast cancer cell lines was evaluated in CCLE database. Meanwhile, HPA database was used to analyze the expression of AGR2 protein in normal and breast cancer tissues. Besides,the gene expression microarray data download from CCLE database was analyzed by using R software to obtain genes co-expressed with AGR2. Functional annotation of AGR2 co-expressed genes was performed by using GO Enrichment and KEGG pathway analyses.Results: Oncomine and GEPIA databases showed that AGR2 gene was highly expressed in breast cancer tissues, and CCLE database analysis showed that AGR2 was highly expressed in all breast cancer cell lines. Immunohistochemistry results from the HPA database showed that the expression of AGR2 protein was significantly higher in breast cancer tissues compared with normal tissues. A total of 946 genes co-expressed with AGR2 in breast cancer were screened out with the R software. With the GO function Enrichment analysis,the co-expressed genes were demonstrated to be mainly involved in biological functions, such as protein localization to cell periphery,protein localization to plasma membrane, cell junction assembly, cell-substrate adhesion, and cell junction organization etc. In addition,the KEGG analysis results showed that co-expressed genes were mainly involved in the progression of gastric cancer and breast cancer,and were associated with proteoglycans in cancer, as well as proline, leucine and isoleucine degradation pathways. Conclusions: AGR2 is highly expressed in breast cancer tissues, which may be a potential oncogenic gene and a new therapeutic target of breast cancer.
2020, 27(3):309-314.
DOI: 10.3872/j.issn.1007-385X.2020.03.015
Abstract:
Objective: To systematically evaluate the efficacy and safety of PD-1/PD-L1 inhibitor combined with chemotherapy as comparing with chemotherapy alone for the first-line treatment of advanced NSCLC (non-small lung cancer). Methods: RCTs (randomized controlled trials) on PD-1/PD-L1 inhibitor combined with chemotherapy compared with chemotherapy alone for the first-line treatment of advanced NSCLC were searched in the PubMed, Cochrane Library, EMbase, EBSCO, Chinese Biomedical Literature Database (CBM), Chinese Journal Full-text Database (CNKI), and Chinese Scientific Journal Full-text Database (VIP). RevMan 5.2 software was used for the Meta-analysis. Results: Six RCTs with 3 238 advanced NSCLC patients were included in this study. Meta-analysis showed that the combination therapy group was more effective than the chemotherapy alone group in OS (HR=0.86, 95%CI=0.79~ 0.94, P=0.0006) and PFS (HR=0.81, 95%CI=0.78~0.84, P<0.00001). The incidence of adverse reactions, such as thrombocytopenia of grade 1-5, vomiting, diarrhea, hypothyroidism, hyperthyroidism, rash, pneumonitis, colitis, hepatitis, dysgeusia, hepatitis of grade 3-5 and colitis, in combined treatment group were all higher than those in chemotherapy alone group, the differences were statistically significant (P<0.01 or P<0.05). Conclusions: Compared with chemotherapy alone, PD-1/PD-L1 inhibitor combined with chemotherapy can significantly improve the OS and PFS of patients with advanced NSCLC in the first-line treatment, while the overall incidence of adverse reactions is higher than chemotherapy.
Objective: To systematically evaluate the efficacy and safety of PD-1/PD-L1 inhibitor combined with chemotherapy as comparing with chemotherapy alone for the first-line treatment of advanced NSCLC (non-small lung cancer). Methods: RCTs (randomized controlled trials) on PD-1/PD-L1 inhibitor combined with chemotherapy compared with chemotherapy alone for the first-line treatment of advanced NSCLC were searched in the PubMed, Cochrane Library, EMbase, EBSCO, Chinese Biomedical Literature Database (CBM), Chinese Journal Full-text Database (CNKI), and Chinese Scientific Journal Full-text Database (VIP). RevMan 5.2 software was used for the Meta-analysis. Results: Six RCTs with 3 238 advanced NSCLC patients were included in this study. Meta-analysis showed that the combination therapy group was more effective than the chemotherapy alone group in OS (HR=0.86, 95%CI=0.79~ 0.94, P=0.0006) and PFS (HR=0.81, 95%CI=0.78~0.84, P<0.00001). The incidence of adverse reactions, such as thrombocytopenia of grade 1-5, vomiting, diarrhea, hypothyroidism, hyperthyroidism, rash, pneumonitis, colitis, hepatitis, dysgeusia, hepatitis of grade 3-5 and colitis, in combined treatment group were all higher than those in chemotherapy alone group, the differences were statistically significant (P<0.01 or P<0.05). Conclusions: Compared with chemotherapy alone, PD-1/PD-L1 inhibitor combined with chemotherapy can significantly improve the OS and PFS of patients with advanced NSCLC in the first-line treatment, while the overall incidence of adverse reactions is higher than chemotherapy.
2020, 27(3):315-320.
DOI: 10.3872/j.issn.1007-385X.2020.03.016
Abstract:
长链非编码RNA（lncRNA）是一类长度大于200 nt、且不编码的RNA。lncRNA 已被证明与人类疾病紧密相关，尤其是肿瘤发生发展。研究表明，肿瘤中一些异常表达的lncRNA 可以通过不同的信号通路，如Wnt/β-catenin 信号通路，促进肿瘤进展过程。在不同肿瘤组织中具有特异性表达特征的lncRNA与Wnt/β-catenin 信号通路之间的相互作用显示出其作为新的生物标志物和治疗靶点的潜能。本文就Wnt/β-catenin 信号通路相关lncRNA 通过调控Wnt/β-catenin 信号转导，影响不同肿瘤类型发生发展的作用进行综述。本文结果或可为临床肿瘤诊断和治疗提供新的思路。
长链非编码RNA（lncRNA）是一类长度大于200 nt、且不编码的RNA。lncRNA 已被证明与人类疾病紧密相关，尤其是肿瘤发生发展。研究表明，肿瘤中一些异常表达的lncRNA 可以通过不同的信号通路，如Wnt/β-catenin 信号通路，促进肿瘤进展过程。在不同肿瘤组织中具有特异性表达特征的lncRNA与Wnt/β-catenin 信号通路之间的相互作用显示出其作为新的生物标志物和治疗靶点的潜能。本文就Wnt/β-catenin 信号通路相关lncRNA 通过调控Wnt/β-catenin 信号转导，影响不同肿瘤类型发生发展的作用进行综述。本文结果或可为临床肿瘤诊断和治疗提供新的思路。
2020, 27(3):321-326.
DOI: 10.3872/j.issn.1007-385X.2020.03.017
Abstract:
染色体外DNA（extrachromosomal DNA，ecDNA）是存在于肿瘤细胞中、可在有丝分裂中期被光镜观察到的游离于染色体外的环形DNA。ecDNA不仅携带高拷贝的癌基因，而且转录活跃，是癌基因表达的主要来源；另外其还是形成肿瘤异质性的关键。本文将就上述ecDNA对肿瘤的作用机制进行概述，并提出关于该领域的研究展望。
染色体外DNA（extrachromosomal DNA，ecDNA）是存在于肿瘤细胞中、可在有丝分裂中期被光镜观察到的游离于染色体外的环形DNA。ecDNA不仅携带高拷贝的癌基因，而且转录活跃，是癌基因表达的主要来源；另外其还是形成肿瘤异质性的关键。本文将就上述ecDNA对肿瘤的作用机制进行概述，并提出关于该领域的研究展望。
2020, 27(3):327-332.
DOI: 10.3872/j.issn.1007-385X.2020.03.018
Abstract:
卵巢上皮癌（epithelial ovarian cancer，EOC）是妇科肿瘤中最致命的恶性肿瘤，在进行最大限度的肿瘤细胞减灭术后需结合铂类药物及紫杉醇联合化疗。多数临床化疗结果表明，EOC患者肿瘤复发后常引起化疗药物的耐药，导致预后差、病死率高。锌指E盒结合同源框1（zinc finger E-box-binding homeobox 1，ZEB1）是多种肿瘤发生、发展、迁移、转移和侵袭的重要调控因子。ZEB1 可能通过调控上皮间质转化（epithelial-mesenchymal transition，EMT）过程、非编码RNA（如miRNA、lncRNA）调控影响卵巢癌耐药；此外，ZEB1 还可介导p73 和BRCA1 相关的表观遗传调控参与卵巢癌的耐药。本文从ZEB1 的结构与生理功能、在卵巢癌发生发展和在卵巢癌耐药中的作用及其可能涉及的机制，以及其作为卵巢癌生物标志物的潜力等方面做一综述，为临床治疗EOC寻找新的治疗策略。
卵巢上皮癌（epithelial ovarian cancer，EOC）是妇科肿瘤中最致命的恶性肿瘤，在进行最大限度的肿瘤细胞减灭术后需结合铂类药物及紫杉醇联合化疗。多数临床化疗结果表明，EOC患者肿瘤复发后常引起化疗药物的耐药，导致预后差、病死率高。锌指E盒结合同源框1（zinc finger E-box-binding homeobox 1，ZEB1）是多种肿瘤发生、发展、迁移、转移和侵袭的重要调控因子。ZEB1 可能通过调控上皮间质转化（epithelial-mesenchymal transition，EMT）过程、非编码RNA（如miRNA、lncRNA）调控影响卵巢癌耐药；此外，ZEB1 还可介导p73 和BRCA1 相关的表观遗传调控参与卵巢癌的耐药。本文从ZEB1 的结构与生理功能、在卵巢癌发生发展和在卵巢癌耐药中的作用及其可能涉及的机制，以及其作为卵巢癌生物标志物的潜力等方面做一综述，为临床治疗EOC寻找新的治疗策略。
2020, 27(3):333-337.
DOI: 10.3872/j.issn.1007-385X.2020.03.019
Abstract:
腹膜转移癌是临床常见难题，其局部微环境近年来已逐步被关注和研究。腹膜和腹水构成了腹膜转移性癌灶特殊的局部微环境，巨噬细胞是腹膜转移癌微环境中占比最多的免疫细胞群体，具有多种亚型，分泌多种细胞因子，参与多条信号通路，在癌细胞存活、血管生成、腹膜侵袭和腹膜转移癌形成的过程中发挥了重要作用。近年来，细胞免疫治疗取得突破性进展，其中靶向肿瘤微环境中肿瘤相关巨噬细胞的免疫治疗逐步开展，多项临床试验正在进行中。本文对巨噬细胞在癌症腹膜转移过程中发挥的作用，以及针对巨噬细胞可能的治疗靶点进行综述。
腹膜转移癌是临床常见难题，其局部微环境近年来已逐步被关注和研究。腹膜和腹水构成了腹膜转移性癌灶特殊的局部微环境，巨噬细胞是腹膜转移癌微环境中占比最多的免疫细胞群体，具有多种亚型，分泌多种细胞因子，参与多条信号通路，在癌细胞存活、血管生成、腹膜侵袭和腹膜转移癌形成的过程中发挥了重要作用。近年来，细胞免疫治疗取得突破性进展，其中靶向肿瘤微环境中肿瘤相关巨噬细胞的免疫治疗逐步开展，多项临床试验正在进行中。本文对巨噬细胞在癌症腹膜转移过程中发挥的作用，以及针对巨噬细胞可能的治疗靶点进行综述。
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- 《中国肿瘤生物治疗杂志》
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Chinese Journal Of Cancer Biotheray ® 2025 All Rights Reserved
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Chinese Journal Of Cancer Biotheray ® 2025 All Rights Reserved
Supported by:Beijing E-Tiller Technology Development Co., Ltd.