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Volume 28,Issue 12,2021 Table of Contents
2021, 28(12):1151-1159.
Abstract:
Objective: To detect the expression level of miR-26b-3p in esophageal squamous cell carcinoma (ESCC) tissues, and to explore its effect on the proliferation, invasion and migration of ESCC cells and its molecular mechanism. Methods: 60 pairs of ESCC tissues and corresponding adjacent tissues that surgically resected from April 1, 2018 to December 25, 2018 were collected from the Fourth Hospital of Hebei Medical University. qPCR was used to detect the expression of miR-26b-3p in ESCC tissues, corresponding adjacent tissues and ESCC cells. ESCC cell lines TE1 and KYSE150 with low miR-26b-3p expression were selected and transfected with miR-26b-3p mimics. At the same time, the negative control was set up. The effects of miR-26b-3p on proliferation, migration and invasion of TE1 and KYSE150 cells were detected by cell proliferation assay, Wound healing assay and Transwell assay. Luciferase reporter gene analysis was used to detect the binding between miR-26b-3p and STAT3 3'UTR. Then, miR-26b-3p mimics and pcDNA3.0-STAT3 were co-transfected into the cells to verify whether STAT3 could reverse the effect of miR-26b-3p on cell proliferation, migration and invasion using cell proliferation assay, Wound healing assay and Transwell test. The effects of methylase inhibitor 5-Aza-DC on the methylation of ESCC cells as well as the expression of miR-26b-3p and STAT3 were detected by qPCR and WB. Results: The expression of miR-26b-3p in ESCC tissues was lower than that in the adjacent normal tissues (P<0.05), and the expression was also significantly lower in ESCC cell lines (TE1, KYSE150 and LYSE170) as compared with normal human esophageal epithelial HEEC cells (all P<0.01). Compared with miR-26b-3p NC group, transfection of miR-26b-3p mimics significantly up[1]regulated the expression of miR-26b-3p in TE1 and KYSE150 cells and obviously inhibited the proliferation, invasion and migration of the cells (P<0.05 or P<0.01). The luciferase reporter gene assay showed that miR-26b-3p significantly inhibited the luciferase activity of wild-type STAT3 vector in TE1 and KYSE150 cells (P<0.05 or P<0.01), while the luciferase activity of mutant-type was not affected. Co-transfection with miR-26b-3p mimics and pcDNA3.0-STAT3 partially reversed the inhibitory effect of miR-26b-3p on proliferation, migration and invasion of TE1 and KYSE150 cells (P<0.05 or P<0.01). After 5-aza-DC treatment, the expression of miR[1]26b-3p was up-regulated (P<0.01), mRNA and protein levels of STAT3 were decreased (P<0.05), and miR-26b-3p was demethylated in TE1 and KYSE150 cells. Conclusion: miR-26b-3p is down-regulated in ESCC tissues and cells due to the hypermethylation of its promoter. As a tumor suppressor, miR-26b-3p inhibits the proliferation, invasion and migration of ESCC cells via targeting STAT3.
Objective: To detect the expression level of miR-26b-3p in esophageal squamous cell carcinoma (ESCC) tissues, and to explore its effect on the proliferation, invasion and migration of ESCC cells and its molecular mechanism. Methods: 60 pairs of ESCC tissues and corresponding adjacent tissues that surgically resected from April 1, 2018 to December 25, 2018 were collected from the Fourth Hospital of Hebei Medical University. qPCR was used to detect the expression of miR-26b-3p in ESCC tissues, corresponding adjacent tissues and ESCC cells. ESCC cell lines TE1 and KYSE150 with low miR-26b-3p expression were selected and transfected with miR-26b-3p mimics. At the same time, the negative control was set up. The effects of miR-26b-3p on proliferation, migration and invasion of TE1 and KYSE150 cells were detected by cell proliferation assay, Wound healing assay and Transwell assay. Luciferase reporter gene analysis was used to detect the binding between miR-26b-3p and STAT3 3'UTR. Then, miR-26b-3p mimics and pcDNA3.0-STAT3 were co-transfected into the cells to verify whether STAT3 could reverse the effect of miR-26b-3p on cell proliferation, migration and invasion using cell proliferation assay, Wound healing assay and Transwell test. The effects of methylase inhibitor 5-Aza-DC on the methylation of ESCC cells as well as the expression of miR-26b-3p and STAT3 were detected by qPCR and WB. Results: The expression of miR-26b-3p in ESCC tissues was lower than that in the adjacent normal tissues (P<0.05), and the expression was also significantly lower in ESCC cell lines (TE1, KYSE150 and LYSE170) as compared with normal human esophageal epithelial HEEC cells (all P<0.01). Compared with miR-26b-3p NC group, transfection of miR-26b-3p mimics significantly up[1]regulated the expression of miR-26b-3p in TE1 and KYSE150 cells and obviously inhibited the proliferation, invasion and migration of the cells (P<0.05 or P<0.01). The luciferase reporter gene assay showed that miR-26b-3p significantly inhibited the luciferase activity of wild-type STAT3 vector in TE1 and KYSE150 cells (P<0.05 or P<0.01), while the luciferase activity of mutant-type was not affected. Co-transfection with miR-26b-3p mimics and pcDNA3.0-STAT3 partially reversed the inhibitory effect of miR-26b-3p on proliferation, migration and invasion of TE1 and KYSE150 cells (P<0.05 or P<0.01). After 5-aza-DC treatment, the expression of miR[1]26b-3p was up-regulated (P<0.01), mRNA and protein levels of STAT3 were decreased (P<0.05), and miR-26b-3p was demethylated in TE1 and KYSE150 cells. Conclusion: miR-26b-3p is down-regulated in ESCC tissues and cells due to the hypermethylation of its promoter. As a tumor suppressor, miR-26b-3p inhibits the proliferation, invasion and migration of ESCC cells via targeting STAT3.
2021, 28(12):1160-1167.
Abstract:
Objective: To explore the expression and clinical significance of lncRNA DNM3OS (dynamin 3 opposite strand/antisense RNA) in laryngeal squamous cell carcinoma (LSCC) tissues and cell lines, and to investigate its effects on in vitro proliferation,migration and invasion of LSCC TU177 cells, as well as to discuss the relationship between DNM3OS and epithelial-mesenchymal transition (EMT). Methods: Sixty-eight pairs of cancer and corresponding para-cancerous tissues from LSCC patients that admitted for surgery from March 2014 to December 2018 were collected from the biological specimen bank of the Fourth hospital of Hebei Medical University. The level of DNM3OS expression in LSCC tissues and cell lines was detected by qPCR. siRNA was used to knockdown DNM3OS expression in TU177 cell. MTS, colony formation and Transwell chamber assays were performed to detect the effect of DNM3OS knockdown on proliferation, migration and invasion of TU177 cells. qPCR and WB methods were used to detect the mRNA and protein expression of EMT-related markers, such as E-cadherin, N-cadherin, vimentin, twist, and SNAI2 after DNM3OS knockdown. Results: The expression level of DNM3OS in LSCC tissues was markedly higher than that in para-cancerous tissues (P<0.01). Upregulated DNM3OS expression was related to TNM stage, lymph node metastasis, and survival of patients with LSCC (P<0.05 or P<0.01). Furthermore, DNM3OS was highly expressed in five LSCC cell lines (Hep-2, AMC-HN-8, TU177, TU212, and TU686) (P<0.05 or P<0.01). The expression of DNM3OS was significantly decreased in TU177 cells after transfection of si-DNM3OS (P<0.01). Compared with the control group, DNM3OS knockdown could suppress the in vitro proliferation, migration and invasion of TU177 cells (all P<0.01), upregulate the expression level of E-cadherin, while down-regulate the expression of N-cadherin,vimentin, twist and SNAI2 in TU177 cells (all P<0.01). Conclusion: DNM3OS overexpression is related to the malignant progression of LSCC, which may be a potential prognostic marker for LSCC patients. DNM3OS may promote the invasion and metastasis of LSCC cells by mediating EMT.
Objective: To explore the expression and clinical significance of lncRNA DNM3OS (dynamin 3 opposite strand/antisense RNA) in laryngeal squamous cell carcinoma (LSCC) tissues and cell lines, and to investigate its effects on in vitro proliferation,migration and invasion of LSCC TU177 cells, as well as to discuss the relationship between DNM3OS and epithelial-mesenchymal transition (EMT). Methods: Sixty-eight pairs of cancer and corresponding para-cancerous tissues from LSCC patients that admitted for surgery from March 2014 to December 2018 were collected from the biological specimen bank of the Fourth hospital of Hebei Medical University. The level of DNM3OS expression in LSCC tissues and cell lines was detected by qPCR. siRNA was used to knockdown DNM3OS expression in TU177 cell. MTS, colony formation and Transwell chamber assays were performed to detect the effect of DNM3OS knockdown on proliferation, migration and invasion of TU177 cells. qPCR and WB methods were used to detect the mRNA and protein expression of EMT-related markers, such as E-cadherin, N-cadherin, vimentin, twist, and SNAI2 after DNM3OS knockdown. Results: The expression level of DNM3OS in LSCC tissues was markedly higher than that in para-cancerous tissues (P<0.01). Upregulated DNM3OS expression was related to TNM stage, lymph node metastasis, and survival of patients with LSCC (P<0.05 or P<0.01). Furthermore, DNM3OS was highly expressed in five LSCC cell lines (Hep-2, AMC-HN-8, TU177, TU212, and TU686) (P<0.05 or P<0.01). The expression of DNM3OS was significantly decreased in TU177 cells after transfection of si-DNM3OS (P<0.01). Compared with the control group, DNM3OS knockdown could suppress the in vitro proliferation, migration and invasion of TU177 cells (all P<0.01), upregulate the expression level of E-cadherin, while down-regulate the expression of N-cadherin,vimentin, twist and SNAI2 in TU177 cells (all P<0.01). Conclusion: DNM3OS overexpression is related to the malignant progression of LSCC, which may be a potential prognostic marker for LSCC patients. DNM3OS may promote the invasion and metastasis of LSCC cells by mediating EMT.
2021, 28(12):1168-1173.
Abstract:
Objective:To explore the role of lncRNA ZNF674-AS1 in cervical cancer and its molecular mechanism. Methods: qPCR was used to detect the expression of ZNF674-AS1 in 31 pairs of cervical cancer tissues and corresponding para-cancerous tissues that obtained from patients who had received surgical treatment during January 2019 and July 2020 in Wuhan Children’s Hospital, as well as in cervical cancer cell lines (SiHa, HeLa, C33A and HCC94) and immortalized cervical epithelial cell lines. The NF674-AS1 over-expression plasmids or the negative control plasmids were transfected into the HCC94 cell line with the least expression of ZNF674-AS1. CCK-8 method and Transwell assay were used to detect the effect of ZNF674-AS1 over-expression on the proliferation and migration of HCC94 cells.Bioinformatics methods and Dual luciferase reporter gene experiment were respectively used to predict and verify the complementary binding relationship among ZNF674-AS1, miR-510-5p and REPS2 (RALBP1 associated Eps domain containing 2). qPCR was conducted to examine the effect of ZNF674-AS1 over-expression on the levels of miR-510-5p and REPS2, and WB assay was used to detect the effect of ZNF674- AS1 over-expression on the level of molecules that related with cell proliferation and migration. Results: Compared with para-cancerous tissues, ZNF674-AS1 was significantly down-regulated in cervical cancer tissues (P<0.01). Compared with immortalized cervical epithelial cells, ZNF674-AS1 was also obviously down-regulated in cervical cancer cell lines (P<0.05 or P<0.01), with the lowest expression in HCC94 cells (P<0.01). Over-expression of ZNF674-AS1 could significantly inhibit the proliferation (P<0.05) and migration (P<0.01) of HCC94 cells. miR-510-5p was proved to interact with ZNF674-AS1, and REPS2 was proved to interact with miR-510-5p. Over-expression of ZNF674-AS1 caused a decrease in the expression level of miR-510-5p (P<0.01). but an increase in the expression level of REPS2 gene (P<0.01) in HCC94 cells, as well as the up- or down- regulation of multiple molecules (CDK2, cyclinD3, vimentin and twist) that related with cell proliferation and migration. Conclusions：lncRNA ZNF674-AS1 is low-expressed in cervical cancer tissues and cell lines and may up-regulate the expression of REPS2 gene by competitively binding miR-510-5p, thereby inhibiting the proliferation and migration of cervical cancer HCC94 cells.
Objective:To explore the role of lncRNA ZNF674-AS1 in cervical cancer and its molecular mechanism. Methods: qPCR was used to detect the expression of ZNF674-AS1 in 31 pairs of cervical cancer tissues and corresponding para-cancerous tissues that obtained from patients who had received surgical treatment during January 2019 and July 2020 in Wuhan Children’s Hospital, as well as in cervical cancer cell lines (SiHa, HeLa, C33A and HCC94) and immortalized cervical epithelial cell lines. The NF674-AS1 over-expression plasmids or the negative control plasmids were transfected into the HCC94 cell line with the least expression of ZNF674-AS1. CCK-8 method and Transwell assay were used to detect the effect of ZNF674-AS1 over-expression on the proliferation and migration of HCC94 cells.Bioinformatics methods and Dual luciferase reporter gene experiment were respectively used to predict and verify the complementary binding relationship among ZNF674-AS1, miR-510-5p and REPS2 (RALBP1 associated Eps domain containing 2). qPCR was conducted to examine the effect of ZNF674-AS1 over-expression on the levels of miR-510-5p and REPS2, and WB assay was used to detect the effect of ZNF674- AS1 over-expression on the level of molecules that related with cell proliferation and migration. Results: Compared with para-cancerous tissues, ZNF674-AS1 was significantly down-regulated in cervical cancer tissues (P<0.01). Compared with immortalized cervical epithelial cells, ZNF674-AS1 was also obviously down-regulated in cervical cancer cell lines (P<0.05 or P<0.01), with the lowest expression in HCC94 cells (P<0.01). Over-expression of ZNF674-AS1 could significantly inhibit the proliferation (P<0.05) and migration (P<0.01) of HCC94 cells. miR-510-5p was proved to interact with ZNF674-AS1, and REPS2 was proved to interact with miR-510-5p. Over-expression of ZNF674-AS1 caused a decrease in the expression level of miR-510-5p (P<0.01). but an increase in the expression level of REPS2 gene (P<0.01) in HCC94 cells, as well as the up- or down- regulation of multiple molecules (CDK2, cyclinD3, vimentin and twist) that related with cell proliferation and migration. Conclusions：lncRNA ZNF674-AS1 is low-expressed in cervical cancer tissues and cell lines and may up-regulate the expression of REPS2 gene by competitively binding miR-510-5p, thereby inhibiting the proliferation and migration of cervical cancer HCC94 cells.
WANG Yang
,
XU Wenguang
,
DONG Yan
,
ZHAO Chunwu
,
XU Youchao
,
ZHAO Haojun
,
SHAN Baoqiang
,
SUN Xiwen
,
SUN Yukai
2021, 28(12):1174-1178.
Abstract:
Objective: To explore the effect of lncRNA-Z38 on malignant biological behavior of gastric cancer cells, and the possible regulatory mechanism. Methods: Gastric cancer cell lines (AGS,MKN74 cells) stably over-expressing lncRNA-Z38 were constructed，and the effect of lncRNA-Z38 over-expression on the stemness of gastric cancer cells was detected through sphere forming assay. High-throughput sequencing of AGS cells with lncRNA-Z38 over-expression and the control cells was performed, and KEGG enrichment analysis was done on the differentially expressed genes to find the potential signaling pathway involved in regulating the occurrence and development of gastric cancer. Furthermore, WB was conducted to verify that lncRNA-Z38 regulates the signaling pathway to promote the stemness of gastric cancer cells. Results: Cell lines with stable lncRNA-Z38 over-expression were successfully constructed. Sphere forming assay suggested that the sphere-forming ability of lncRNA-Z38 overexpressed gastric cancer cells was significantly higher than their control cells (P<0.05). According to the high-throughput sequencing results, 1 999 differentially expressed genes in lncRNA-Z38 over-expressed gastric cancers cells were screened out, among which 1 238 were up-regulated and 761 were downregulated. KEGG pathway enrichment analysis showed that the most significantly enriched pathway was TGF-β signaling pathway (P<0.05). Further, WB results verified that the encoding proteins of TGF-β pathway constituent genes ID3 and BMP6 were significantly up-regulated while GDF-5 and WNT8B were significantly down-regulated in lncRNA-Z38 over-expressed gastric cancer cells (P<0.05). Conclusion: Highly expressed lncRNA-Z38 enhances the stemness of gastric cancer cells through regulating the TGF-β pathway.
Objective: To explore the effect of lncRNA-Z38 on malignant biological behavior of gastric cancer cells, and the possible regulatory mechanism. Methods: Gastric cancer cell lines (AGS,MKN74 cells) stably over-expressing lncRNA-Z38 were constructed，and the effect of lncRNA-Z38 over-expression on the stemness of gastric cancer cells was detected through sphere forming assay. High-throughput sequencing of AGS cells with lncRNA-Z38 over-expression and the control cells was performed, and KEGG enrichment analysis was done on the differentially expressed genes to find the potential signaling pathway involved in regulating the occurrence and development of gastric cancer. Furthermore, WB was conducted to verify that lncRNA-Z38 regulates the signaling pathway to promote the stemness of gastric cancer cells. Results: Cell lines with stable lncRNA-Z38 over-expression were successfully constructed. Sphere forming assay suggested that the sphere-forming ability of lncRNA-Z38 overexpressed gastric cancer cells was significantly higher than their control cells (P<0.05). According to the high-throughput sequencing results, 1 999 differentially expressed genes in lncRNA-Z38 over-expressed gastric cancers cells were screened out, among which 1 238 were up-regulated and 761 were downregulated. KEGG pathway enrichment analysis showed that the most significantly enriched pathway was TGF-β signaling pathway (P<0.05). Further, WB results verified that the encoding proteins of TGF-β pathway constituent genes ID3 and BMP6 were significantly up-regulated while GDF-5 and WNT8B were significantly down-regulated in lncRNA-Z38 over-expressed gastric cancer cells (P<0.05). Conclusion: Highly expressed lncRNA-Z38 enhances the stemness of gastric cancer cells through regulating the TGF-β pathway.
2021, 28(12):1179-1185.
Abstract:
Objective: To evaluate the treatment efficacy of mucin 1 (MUC1) gene transfected dendritic cell (MUC1-DC) vaccine on breast cancer MCF-7 cell xenografts in nude mice and its possible mechanism. Methods: GFP lentivirus were transfected into human breast cancer MCF-7 cells to obtain the GFP-MCF-7 cells which were subcutaneously implanted into BALB/c nude mice. After tumorigenesis, the mice were randomly divided into three groups. First, CIK cells activated in vitro (1×108 cells/mouse) were injected into the mice of each group via tail vein. In the treatment groups, MUC1-DC (MUC1-DC group) or DC (DC group) were injected subcutaneously (0.2 ml, 1×107 cells/mouse). In the control group, normal saline of the same volume was injected. The treatment frequency was once per day for 5 consecutive days. The fluorescence imaging of transplanted tumor was observed before and 35 days after the treatment with optical imaging system, and fluorescence intensity and area were analyzed. The expression of Caspase 3 in xenograft tissues was detected by immunohistochemistry, and the cell apoptosis was detected by TUNEL assay. Results: The tumor formation rate of nude mice was 100% at 7 days after GFP-MCF-7 implantation. The results of optical molecular imaging showed that there was no significant difference in fluorescence signal intensity among MUC1-DC group, DC group and Control group before treatment (P>0.05). At day 35 after treatment, the fluorescence signal intensity of MUC1-DC group and DC group was significantly lower than that of Control group (P<0.05). There was no significant difference between DC and Control group, as well as between MUC1-DC and DC group (all P>0.05), but the fluorescence signal of MUC1-DC group was lower than that of DC group. There was no significant difference in the distribution area of fluorescence signal among MUC1-DC group, DC group and Control group before treatment (P>0.05). At day 35, the fluorescence signal in Control group was scattered in multiple areas, and the area of fluorescence signal in MUC1-DC group and DC group was significantly smaller than that in Control group (all P<0.01), but there was no significant difference between MUC1-DC group and DC group (P>0.05). Caspase 3 expression was the highest in MUC1-DC group, followed by DC group, and the least in Control group, and the differences among the three groups were significant (all P<0.05). TUNEL results showed that the apoptosis rate of Control group, DC group and MUC1-DC group was (4.11±2.61)% , (9.63±2.27)% , and (25.30±8.24)% , respectively (all P<0.05). Conclusion: Compared with mere DC immunotherapy, MUC1-DC vaccine can more effectively inhibit tumor growth and metastasis in nude mice bearing human breast cancer cell xenografts, exerting a better effect in promoting tumor cell apoptosis.
Objective: To evaluate the treatment efficacy of mucin 1 (MUC1) gene transfected dendritic cell (MUC1-DC) vaccine on breast cancer MCF-7 cell xenografts in nude mice and its possible mechanism. Methods: GFP lentivirus were transfected into human breast cancer MCF-7 cells to obtain the GFP-MCF-7 cells which were subcutaneously implanted into BALB/c nude mice. After tumorigenesis, the mice were randomly divided into three groups. First, CIK cells activated in vitro (1×108 cells/mouse) were injected into the mice of each group via tail vein. In the treatment groups, MUC1-DC (MUC1-DC group) or DC (DC group) were injected subcutaneously (0.2 ml, 1×107 cells/mouse). In the control group, normal saline of the same volume was injected. The treatment frequency was once per day for 5 consecutive days. The fluorescence imaging of transplanted tumor was observed before and 35 days after the treatment with optical imaging system, and fluorescence intensity and area were analyzed. The expression of Caspase 3 in xenograft tissues was detected by immunohistochemistry, and the cell apoptosis was detected by TUNEL assay. Results: The tumor formation rate of nude mice was 100% at 7 days after GFP-MCF-7 implantation. The results of optical molecular imaging showed that there was no significant difference in fluorescence signal intensity among MUC1-DC group, DC group and Control group before treatment (P>0.05). At day 35 after treatment, the fluorescence signal intensity of MUC1-DC group and DC group was significantly lower than that of Control group (P<0.05). There was no significant difference between DC and Control group, as well as between MUC1-DC and DC group (all P>0.05), but the fluorescence signal of MUC1-DC group was lower than that of DC group. There was no significant difference in the distribution area of fluorescence signal among MUC1-DC group, DC group and Control group before treatment (P>0.05). At day 35, the fluorescence signal in Control group was scattered in multiple areas, and the area of fluorescence signal in MUC1-DC group and DC group was significantly smaller than that in Control group (all P<0.01), but there was no significant difference between MUC1-DC group and DC group (P>0.05). Caspase 3 expression was the highest in MUC1-DC group, followed by DC group, and the least in Control group, and the differences among the three groups were significant (all P<0.05). TUNEL results showed that the apoptosis rate of Control group, DC group and MUC1-DC group was (4.11±2.61)% , (9.63±2.27)% , and (25.30±8.24)% , respectively (all P<0.05). Conclusion: Compared with mere DC immunotherapy, MUC1-DC vaccine can more effectively inhibit tumor growth and metastasis in nude mice bearing human breast cancer cell xenografts, exerting a better effect in promoting tumor cell apoptosis.
2021, 28(12):1186-1193.
Abstract:
Objective: To screen the key differentially expressed miRNAs and their target genes in breast cancer using bioinformatics tools, and to observe the effect of interfering their expression on the function of breast cancer cells. Methods: The GEO database was used to screen the differentially expressed miRNAs in breast cancer, and the ENCOR1 database was further used to verify the expression of screened miRNAs to identify the most differentially expressed miRNA as the research target. Starbase, miRDB and miRWalk databases were used to predict the target genes of miR-32-5p. GO analysis and KEGG analysis of target genes were done by using DAVID database. PPI network analysis and screening of hub genes were performed using String database and Cytoscape3.6.2 software. The Dickkopf-related protein 3 (DKK3) gene with the most significant degree was selected from the hub genes for follow[1]up experiments. qPCR was used to detect the expression of miR-32-5p in human normal breast MCF10A cells and human breast cancer cells (MCF7, MDA-MB-231 and MDA-MB-453 cells). MiR-32-5p mimics, miR-32-5p inhibitors and their control (NC) sequences were transfected into MDA-MB-231 cells. The effects of over-expression or inhibition of miR-32-5p on cell proliferation,apoptosis and invasion were detected by CCK-8 method, flow cytometry, and Transwell experiment, respectively. Results: Two differentially expressed miRNAs were identified from the two datasets in GEO database. ENCORI database was adopted to verify the differentially expressed miRNAs and showed that the expression level of miR-32-5p was consistent with the result in the GEO database, so it was selected for subsequent research. 198 potential target genes of miR-32-5p were predicted and 10 hub genes (DKK3,WNT2B, SFRP5, SFRP2, SFRP1, LRP6, WNT6, KREMEN1, NEDD4L and TRIP12) were identified, among which DKK3 showed the most significant degree. So, miR-32-5p/DKK3 axis was selected for the subsequent research. The expression of miR-32-5p in three breast cancer was significantly higher than that in normal mammary gland cells (all P<0.01), with the highest expression in MDA[1]MB-231 cells. Dual-luciferase reporter gene assay verified cell lines that miR-32-5p tageted and down-regulated. After transfection with miR-32-5p mimics or miR-32-5p inhibitors, the expression of miR-32-5p in MDA-MB-231 cells was successfully increased or inhibited. Compared with the control group, overexpression of miR-32-5p could inhibit the apoptosis of MDA-MB-231 cells and promote cell proliferation and invasion (P<0.05 or P<0.01), while knocking down miR-32-5p played an opposite role (all P<0.01).Conclusion: miR-32-5p/DKK3 axis may be a key pathway affecting the development of breast cancer. Over-expression of miR-32-5p can inhibit the apoptosis but promote cell proliferation and invasion of breast cancer MDA-MB-231 cells.
Objective: To screen the key differentially expressed miRNAs and their target genes in breast cancer using bioinformatics tools, and to observe the effect of interfering their expression on the function of breast cancer cells. Methods: The GEO database was used to screen the differentially expressed miRNAs in breast cancer, and the ENCOR1 database was further used to verify the expression of screened miRNAs to identify the most differentially expressed miRNA as the research target. Starbase, miRDB and miRWalk databases were used to predict the target genes of miR-32-5p. GO analysis and KEGG analysis of target genes were done by using DAVID database. PPI network analysis and screening of hub genes were performed using String database and Cytoscape3.6.2 software. The Dickkopf-related protein 3 (DKK3) gene with the most significant degree was selected from the hub genes for follow[1]up experiments. qPCR was used to detect the expression of miR-32-5p in human normal breast MCF10A cells and human breast cancer cells (MCF7, MDA-MB-231 and MDA-MB-453 cells). MiR-32-5p mimics, miR-32-5p inhibitors and their control (NC) sequences were transfected into MDA-MB-231 cells. The effects of over-expression or inhibition of miR-32-5p on cell proliferation,apoptosis and invasion were detected by CCK-8 method, flow cytometry, and Transwell experiment, respectively. Results: Two differentially expressed miRNAs were identified from the two datasets in GEO database. ENCORI database was adopted to verify the differentially expressed miRNAs and showed that the expression level of miR-32-5p was consistent with the result in the GEO database, so it was selected for subsequent research. 198 potential target genes of miR-32-5p were predicted and 10 hub genes (DKK3,WNT2B, SFRP5, SFRP2, SFRP1, LRP6, WNT6, KREMEN1, NEDD4L and TRIP12) were identified, among which DKK3 showed the most significant degree. So, miR-32-5p/DKK3 axis was selected for the subsequent research. The expression of miR-32-5p in three breast cancer was significantly higher than that in normal mammary gland cells (all P<0.01), with the highest expression in MDA[1]MB-231 cells. Dual-luciferase reporter gene assay verified cell lines that miR-32-5p tageted and down-regulated. After transfection with miR-32-5p mimics or miR-32-5p inhibitors, the expression of miR-32-5p in MDA-MB-231 cells was successfully increased or inhibited. Compared with the control group, overexpression of miR-32-5p could inhibit the apoptosis of MDA-MB-231 cells and promote cell proliferation and invasion (P<0.05 or P<0.01), while knocking down miR-32-5p played an opposite role (all P<0.01).Conclusion: miR-32-5p/DKK3 axis may be a key pathway affecting the development of breast cancer. Over-expression of miR-32-5p can inhibit the apoptosis but promote cell proliferation and invasion of breast cancer MDA-MB-231 cells.
MAO Lili
,
BAI Xue
,
DAI Jie
,
CUI Chuanliang
,
CHI Zhihong
,
TANG Bixia
,
KONG Yan
,
LIAN Bin
,
WANG Xuan
,
WEI Xiaoting
,
LI Caili
,
GUO Jun
,
SI Lu
2021, 28(12):1194-1200.
Abstract:
Objective: To evaluate the treatment efficacy and safety profile of NCA regimen (nab-paclitaxel, carboplatin and antiangiogenic drug) in advanced melanoma patients who failed previous therapy. Methods: We retrospectively enrolled the melanoma patients who were treated in the Department of Melanoma of Peking University Cancer Hospital from April 1, 2012 to March 31, 2019 and investigated the efficacy and safety of NCA regimen in unresectable stage IIIC or IV melanoma patients who failed previous therapy. The primary endpoint was the progression-free survival (PFS), and the secondary indicators were objective response rate (ORR), overall survival (OS), disease control rate (DCR) and adverse reactions. The patients were divided into endostatint treatment group (Endo-arm, n=73) and bevacizumab treatment group (Beva-arm, n=103) according to the antiangiogenic drug used in the regimen. Propensity score matching was applied to balance the differences in baseline covariates between different antiangiogenic drug groups. Results: Overall, 176 patients were included in this study. The median age of included patients was 51 years old (18-78). Stage IV patients accounted for 97%. There were 50% patients with higher LDH level than normal, and 28% patients with liver metastasis. The proportions of lines of previous treatment were 57% for first line treatment, 33% for second line and 10% for third-fourth line. The median PFS was 3.8 months (95% CI: 3.0–4.6 m) and median OS was 10.5 months (95% CI: 8.9-12.1 m) in overall study population. There were 9 PR (partial remission) cases and 2 CR (complete remission) cases, yielding an overall ORR of 6% with a DCR of 70%. The median PFS of Endo-arm and Beva-arm was 4.7 months (95%CI:3.5~5.9m) and 3.4 months (95%CI: 3.0~4.6m) respectively, while the median OS was 12.2 months (95%CI: 11.1~13.2m) and 9.1 months (95%CI: 7.8~10.4m) respectively. After the age, gender, lines of previous treatment and LDH level of patients were adjusted by propensity score matching, there were no statistical differences in PFS and OS between Beva-arm and Endo-arm. Common adverse events included alopecia, peripheral neuropathy, neutropenia, fatigue and nausea. 26 patients (15%) discontinued treatment adverse events. Conclusion: The NCA regimen shows modest antitumor effects in melanoma patients who failed previous treatment, due to and the adverse reactions were acceptable.
Objective: To evaluate the treatment efficacy and safety profile of NCA regimen (nab-paclitaxel, carboplatin and antiangiogenic drug) in advanced melanoma patients who failed previous therapy. Methods: We retrospectively enrolled the melanoma patients who were treated in the Department of Melanoma of Peking University Cancer Hospital from April 1, 2012 to March 31, 2019 and investigated the efficacy and safety of NCA regimen in unresectable stage IIIC or IV melanoma patients who failed previous therapy. The primary endpoint was the progression-free survival (PFS), and the secondary indicators were objective response rate (ORR), overall survival (OS), disease control rate (DCR) and adverse reactions. The patients were divided into endostatint treatment group (Endo-arm, n=73) and bevacizumab treatment group (Beva-arm, n=103) according to the antiangiogenic drug used in the regimen. Propensity score matching was applied to balance the differences in baseline covariates between different antiangiogenic drug groups. Results: Overall, 176 patients were included in this study. The median age of included patients was 51 years old (18-78). Stage IV patients accounted for 97%. There were 50% patients with higher LDH level than normal, and 28% patients with liver metastasis. The proportions of lines of previous treatment were 57% for first line treatment, 33% for second line and 10% for third-fourth line. The median PFS was 3.8 months (95% CI: 3.0–4.6 m) and median OS was 10.5 months (95% CI: 8.9-12.1 m) in overall study population. There were 9 PR (partial remission) cases and 2 CR (complete remission) cases, yielding an overall ORR of 6% with a DCR of 70%. The median PFS of Endo-arm and Beva-arm was 4.7 months (95%CI:3.5~5.9m) and 3.4 months (95%CI: 3.0~4.6m) respectively, while the median OS was 12.2 months (95%CI: 11.1~13.2m) and 9.1 months (95%CI: 7.8~10.4m) respectively. After the age, gender, lines of previous treatment and LDH level of patients were adjusted by propensity score matching, there were no statistical differences in PFS and OS between Beva-arm and Endo-arm. Common adverse events included alopecia, peripheral neuropathy, neutropenia, fatigue and nausea. 26 patients (15%) discontinued treatment adverse events. Conclusion: The NCA regimen shows modest antitumor effects in melanoma patients who failed previous treatment, due to and the adverse reactions were acceptable.
2021, 28(12):1201-1206.
Abstract:
Objective:To detect the expression of programmed death molecule 1 (PD-1) on T cell surface, the levels of T cell subsets and other immune indices in patients with nasopharyngeal carcinoma (NPC) during chemoradiotherapy, and to discuss the immune function of patients in different treatment periods. Methods: Blood samples of 30 patients who were pathologically confirmed of nasopharyngeal carcinoma in Fujian Cancer Hospital from April 2015 to September 2015 were collected for this study. Flow cytometry was used to dynamically detect the immune indicators of NPC patients in different treatment periods (pre-treatment, post-chemotherapy and post-radiotherapy), including PD-1 on T cell surface, T cell subsets, NK cells and B cells etc. Results: Compared with pre[1]treatment, the proportion of CD3+ T cells, CD4+T cells, CD8+CD28+T cells and CD4+/CD8+ ratio in peripheral blood of patients increased significantly, while the proportion of B cells (CD19+) and NK cells (CD3-CD16+CD56+ cells) decreased significantly after neoadjuvant chemotherapy (all P<0.05). After chemoradiotherapy, the proportion of CD4+T cells, CD8+CD28+T cells and CD4+/CD8+ ratio in peripheral blood of patients decreased to a lower level compared to pre-treatment (all P<0.05), while the proportion of NK cells was up-regulated (P<0.05). Compared with pre-treatment, the expression level of PD-1 on T cells increased significantly after radiotherapy (P<0.05). Conclusion: The study found that the proportion of T cell subsets in peripheral blood increased after neoadjuvant chemotherapy but decreased significantly at the end of radiotherapy, indicating that the immune function of NPC patients was significantly suppressed at the end of radiotherapy. The expression level of PD-1 on T cells was significantly up-regulated after radiotherapy, indicating that the end of chemoradiotherapy maybe an optimal timepoint for anti-PD-1 immunotherapy, which may exert more efficient and long-lasting anti-tumor effect for NPC.
Objective:To detect the expression of programmed death molecule 1 (PD-1) on T cell surface, the levels of T cell subsets and other immune indices in patients with nasopharyngeal carcinoma (NPC) during chemoradiotherapy, and to discuss the immune function of patients in different treatment periods. Methods: Blood samples of 30 patients who were pathologically confirmed of nasopharyngeal carcinoma in Fujian Cancer Hospital from April 2015 to September 2015 were collected for this study. Flow cytometry was used to dynamically detect the immune indicators of NPC patients in different treatment periods (pre-treatment, post-chemotherapy and post-radiotherapy), including PD-1 on T cell surface, T cell subsets, NK cells and B cells etc. Results: Compared with pre[1]treatment, the proportion of CD3+ T cells, CD4+T cells, CD8+CD28+T cells and CD4+/CD8+ ratio in peripheral blood of patients increased significantly, while the proportion of B cells (CD19+) and NK cells (CD3-CD16+CD56+ cells) decreased significantly after neoadjuvant chemotherapy (all P<0.05). After chemoradiotherapy, the proportion of CD4+T cells, CD8+CD28+T cells and CD4+/CD8+ ratio in peripheral blood of patients decreased to a lower level compared to pre-treatment (all P<0.05), while the proportion of NK cells was up-regulated (P<0.05). Compared with pre-treatment, the expression level of PD-1 on T cells increased significantly after radiotherapy (P<0.05). Conclusion: The study found that the proportion of T cell subsets in peripheral blood increased after neoadjuvant chemotherapy but decreased significantly at the end of radiotherapy, indicating that the immune function of NPC patients was significantly suppressed at the end of radiotherapy. The expression level of PD-1 on T cells was significantly up-regulated after radiotherapy, indicating that the end of chemoradiotherapy maybe an optimal timepoint for anti-PD-1 immunotherapy, which may exert more efficient and long-lasting anti-tumor effect for NPC.
2021, 28(12):1207-1214.
Abstract:
食管癌的发病率和死亡率均处于较高水平，严重威胁人类健康，其传统疗法的疗效较差，且目前尚无针对食管癌的特 异性靶向药应用于临床。作为一种新的有效的癌症治疗手段，免疫治疗在食管癌治疗中具有广阔的应用前景。免疫治疗主要包 括免疫检查点抑制剂（immune checkpoint inhibitor，ICI）、过继免疫细胞疗法、联合疗法和肿瘤疫苗疗法等。本文就免疫检查点抑 制剂等免疫治疗方法在食管癌各线治疗中的临床和基础研究进行综述，并探讨食管癌相关生物标志物在肿瘤免疫治疗中的预测 价值。
食管癌的发病率和死亡率均处于较高水平，严重威胁人类健康，其传统疗法的疗效较差，且目前尚无针对食管癌的特 异性靶向药应用于临床。作为一种新的有效的癌症治疗手段，免疫治疗在食管癌治疗中具有广阔的应用前景。免疫治疗主要包 括免疫检查点抑制剂（immune checkpoint inhibitor，ICI）、过继免疫细胞疗法、联合疗法和肿瘤疫苗疗法等。本文就免疫检查点抑 制剂等免疫治疗方法在食管癌各线治疗中的临床和基础研究进行综述，并探讨食管癌相关生物标志物在肿瘤免疫治疗中的预测 价值。
2021, 28(12):1215-1218.
Abstract:
胆固醇是机体细胞膜性结构的重要组分和合成甾体类激素的主要原料，肝脏是机体胆固醇代谢的核心器官，司职胆 固醇合成、摄取、转运和排泄等重要功能。肝脏胆固醇代谢的失调参与一系列肝脏疾病的发生发展，尤其与肝癌的发生发展密切 相关。因此，探索肝脏尤其是肝细胞中胆固醇代谢的分子调控机制，阐明胆固醇在肝癌中的作用，并寻求干预肝脏胆固醇代谢的 潜在靶点，是目前广受关注的重要科学问题。本文就肝脏胆固醇代谢调控参与肝癌发生发展及其生物治疗方面的研究进展作一 综述。
胆固醇是机体细胞膜性结构的重要组分和合成甾体类激素的主要原料，肝脏是机体胆固醇代谢的核心器官，司职胆 固醇合成、摄取、转运和排泄等重要功能。肝脏胆固醇代谢的失调参与一系列肝脏疾病的发生发展，尤其与肝癌的发生发展密切 相关。因此，探索肝脏尤其是肝细胞中胆固醇代谢的分子调控机制，阐明胆固醇在肝癌中的作用，并寻求干预肝脏胆固醇代谢的 潜在靶点，是目前广受关注的重要科学问题。本文就肝脏胆固醇代谢调控参与肝癌发生发展及其生物治疗方面的研究进展作一 综述。
2021, 28(12):1219-1226.
Abstract:
B7/CD28家族分子作为主要的共信号分子，在T细胞功能调控及免疫应答中发挥着至关重要的作用，关于其功能的 研究及应用在世界范围内广泛开展。其中，CD28和CTLA-4都可以与B7-1/B7-2结合，但CD28能够促进T细胞增殖，维持Treg 细胞稳态；而CTLA-4则可以抑制T细胞增殖，影响CD4+ T细胞的分化；抗CTLA-4单抗ipilimumab与抗PD-1单抗联用能够用于 治疗PD-L1+ 的非小细胞肺癌以及无症状的Ⅳ期黑色素瘤。PD-L1/PD-L2：PD-1途径则可以通过多种方式调节T细胞功能，参与 CD8+ T细胞“耗竭”状态的形成与维持。目前，针对PD-L1/PD-L2：PD-1 途径的免疫治疗相关药物开发广泛且较为成熟，抗PD1单抗主要通过诱导肿瘤浸润的部分耗竭的CD8+ T细胞亚群的扩增发挥抗肿瘤作用。ICOS：ICOSL途径在T细胞分化、细胞因 子分泌以及体液免疫应答中起着重要作用，抗ICOS抗体药物feladilimab与抗PD-1单抗联用能够有效治疗多发性或难治性骨髓 瘤。B7-H3同时具有共刺激效应和共抑制效应，阻断B7-H3后能够有效增强TIL的抗肿瘤效应，其单抗enoblituzumab能够与抗 PD-1 单抗联用治疗非小细胞肺癌。B7-H4、人内源性逆转录病毒?H 长末端重复关联蛋白（human endogenous retrovirus?H long terminal repeat?associating protein，HHLA）以及含V结构域抑制T细胞活化的免疫球蛋白（V?domain Ig?cotaining suppressor of T cell activation，VISTA）均能够提供抑制信号，针对B7-H4靶点的CAR-T治疗以及VISTA的小分子抑制剂CA-170均有临床试验 正在进行中。目前，对共信号分子的研究已经获得了长足进展，针对某些活化性分子或者抑制性分子开发了相关抗肿瘤药物并 在临床中取得一定成效，但如何进一步提高其治疗有效率、减少不良反应等仍有待探索。
B7/CD28家族分子作为主要的共信号分子，在T细胞功能调控及免疫应答中发挥着至关重要的作用，关于其功能的 研究及应用在世界范围内广泛开展。其中，CD28和CTLA-4都可以与B7-1/B7-2结合，但CD28能够促进T细胞增殖，维持Treg 细胞稳态；而CTLA-4则可以抑制T细胞增殖，影响CD4+ T细胞的分化；抗CTLA-4单抗ipilimumab与抗PD-1单抗联用能够用于 治疗PD-L1+ 的非小细胞肺癌以及无症状的Ⅳ期黑色素瘤。PD-L1/PD-L2：PD-1途径则可以通过多种方式调节T细胞功能，参与 CD8+ T细胞“耗竭”状态的形成与维持。目前，针对PD-L1/PD-L2：PD-1 途径的免疫治疗相关药物开发广泛且较为成熟，抗PD1单抗主要通过诱导肿瘤浸润的部分耗竭的CD8+ T细胞亚群的扩增发挥抗肿瘤作用。ICOS：ICOSL途径在T细胞分化、细胞因 子分泌以及体液免疫应答中起着重要作用，抗ICOS抗体药物feladilimab与抗PD-1单抗联用能够有效治疗多发性或难治性骨髓 瘤。B7-H3同时具有共刺激效应和共抑制效应，阻断B7-H3后能够有效增强TIL的抗肿瘤效应，其单抗enoblituzumab能够与抗 PD-1 单抗联用治疗非小细胞肺癌。B7-H4、人内源性逆转录病毒?H 长末端重复关联蛋白（human endogenous retrovirus?H long terminal repeat?associating protein，HHLA）以及含V结构域抑制T细胞活化的免疫球蛋白（V?domain Ig?cotaining suppressor of T cell activation，VISTA）均能够提供抑制信号，针对B7-H4靶点的CAR-T治疗以及VISTA的小分子抑制剂CA-170均有临床试验 正在进行中。目前，对共信号分子的研究已经获得了长足进展，针对某些活化性分子或者抑制性分子开发了相关抗肿瘤药物并 在临床中取得一定成效，但如何进一步提高其治疗有效率、减少不良反应等仍有待探索。
2021, 28(12):1227-1231.
Abstract:
外泌体和EMT在宫颈癌发生发展中均发挥了重要作用。外泌体可通过改变肿瘤微环境、促进肿瘤血管生成而影响 宫颈癌疾病进展。EMT可影响肿瘤侵袭迁移能力，宫颈癌细胞的EMT可被miRNA、lncRNA、蛋白质等活性物质调控。而各种细 胞、组织来源的外泌体包含丰富的蛋白质、脂质、核酸等多种活性物质，可直接或通过Wnt/β-catenin、PTEN/P I3K/Akt 等信号通 路调控肿瘤 EMT，然而对外泌体在宫颈癌 EMT 中作用的了解仍有限，还需要大量研究数据支持。对宫颈癌EMT及外泌 体对其的作用展开讨论，拟为研究外泌体在宫颈癌EMT中的作用提供方向，为宫颈癌诊断、治疗等研究提供新的思路。
外泌体和EMT在宫颈癌发生发展中均发挥了重要作用。外泌体可通过改变肿瘤微环境、促进肿瘤血管生成而影响 宫颈癌疾病进展。EMT可影响肿瘤侵袭迁移能力，宫颈癌细胞的EMT可被miRNA、lncRNA、蛋白质等活性物质调控。而各种细 胞、组织来源的外泌体包含丰富的蛋白质、脂质、核酸等多种活性物质，可直接或通过Wnt/β-catenin、PTEN/P I3K/Akt 等信号通 路调控肿瘤 EMT，然而对外泌体在宫颈癌 EMT 中作用的了解仍有限，还需要大量研究数据支持。对宫颈癌EMT及外泌 体对其的作用展开讨论，拟为研究外泌体在宫颈癌EMT中的作用提供方向，为宫颈癌诊断、治疗等研究提供新的思路。
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Chinese Journal Of Cancer Biotheray ® 2025 All Rights Reserved
Supported by:Beijing E-Tiller Technology Development Co., Ltd.