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Volume 28,Issue 10,2021 Table of Contents
2021, 28(10):961-968.
Abstract:
Objective: To investigate the effect and mechanism of lncRNA HOTTIP on proliferation, apoptosis and EMT of lung cancer cells. Methods: The expressions of lncRNA HOTTIP, miR-637 and KLK4 in SPC-A-1, BEAS-2B cells were detected by qPCR.After siRNA interference with the expression of lncRNA HOTTIP, the proliferation, invasion, apoptosis and EMT of SPC-A-1 cells were detected by CCK-8, Transwell, flow cytometry, and WB, respectively. The targeting relationship between lncRNA HOTTIP and miR-637 was analyzed by miRanda software and dual-luciferase reporter gene assay. RNA pull-down assay was used to detect the adsorption of lncRNA HOTTIP and miR-637, and to detect the effects of lncRNA HOTTIP regulating miR-637 on proliferation,invasion, apoptosis, and EMT of SPC-A-1 cells. The correlation between miR-637 and KLK4 was analyzed by TargetScan software,and the interaction between miR-637 and KLK4 was detected by dual-luciferase reporter gene assay. After siRNA interference with the expression of KLK4, the proliferation, invasion, apoptosis, and EMT of SPC-A-1 cells were detected. After down regulation of lncRNA HOTTIP and miR-637 expression, the levels of KLK4 mRNA and protein expression were detected by qPCR and WB. Results:Compared with BEAS-2B cells, the expression of lncRNA HOTTIP in SPC-A-1 cells was significantly up-regulated (P<0.01), the expression of miR-637 was down-regulated (P<0.01), the KLK4 expression was up-regulated (P<0.01). Down-regulation of lncRNA HOTTIP could significantly reduce the proliferation, invasion, and EMT capacity of SPC-A-1 cells, and increase the apoptosis rate (P<0.01). lncRNA HOTTIP had a targeting relationship with miR-637. Down-regulation of miR-637 expression could significantly promote the proliferation, invasion and EMT capacity of SPC-A-1 cells, and inhibit the apoptosis rate (P<0.01). miR-637 specifically bound to KLK4 3'UTR. Down-regulation of KLK4 could significantly inhibit the proliferation, invasion, and EMT capacity of SPC-A-1 cells, and increase the apoptosis rate (P<0.01). Down-regulation of lncRNA HOTTIP could significantly decrease KLK4 expression,while down-regulation of miR-637 could promote KLK4 expression (P<0.05). Conclusion: Up-regulation of lncRNA HOTTIP promotes proliferation, invasion, and EMT of lung cancer SPC-A-1 cells through miR-637/KLK4 axis, and inhibits the apoptosis of cancer cells.
Objective: To investigate the effect and mechanism of lncRNA HOTTIP on proliferation, apoptosis and EMT of lung cancer cells. Methods: The expressions of lncRNA HOTTIP, miR-637 and KLK4 in SPC-A-1, BEAS-2B cells were detected by qPCR.After siRNA interference with the expression of lncRNA HOTTIP, the proliferation, invasion, apoptosis and EMT of SPC-A-1 cells were detected by CCK-8, Transwell, flow cytometry, and WB, respectively. The targeting relationship between lncRNA HOTTIP and miR-637 was analyzed by miRanda software and dual-luciferase reporter gene assay. RNA pull-down assay was used to detect the adsorption of lncRNA HOTTIP and miR-637, and to detect the effects of lncRNA HOTTIP regulating miR-637 on proliferation,invasion, apoptosis, and EMT of SPC-A-1 cells. The correlation between miR-637 and KLK4 was analyzed by TargetScan software,and the interaction between miR-637 and KLK4 was detected by dual-luciferase reporter gene assay. After siRNA interference with the expression of KLK4, the proliferation, invasion, apoptosis, and EMT of SPC-A-1 cells were detected. After down regulation of lncRNA HOTTIP and miR-637 expression, the levels of KLK4 mRNA and protein expression were detected by qPCR and WB. Results:Compared with BEAS-2B cells, the expression of lncRNA HOTTIP in SPC-A-1 cells was significantly up-regulated (P<0.01), the expression of miR-637 was down-regulated (P<0.01), the KLK4 expression was up-regulated (P<0.01). Down-regulation of lncRNA HOTTIP could significantly reduce the proliferation, invasion, and EMT capacity of SPC-A-1 cells, and increase the apoptosis rate (P<0.01). lncRNA HOTTIP had a targeting relationship with miR-637. Down-regulation of miR-637 expression could significantly promote the proliferation, invasion and EMT capacity of SPC-A-1 cells, and inhibit the apoptosis rate (P<0.01). miR-637 specifically bound to KLK4 3'UTR. Down-regulation of KLK4 could significantly inhibit the proliferation, invasion, and EMT capacity of SPC-A-1 cells, and increase the apoptosis rate (P<0.01). Down-regulation of lncRNA HOTTIP could significantly decrease KLK4 expression,while down-regulation of miR-637 could promote KLK4 expression (P<0.05). Conclusion: Up-regulation of lncRNA HOTTIP promotes proliferation, invasion, and EMT of lung cancer SPC-A-1 cells through miR-637/KLK4 axis, and inhibits the apoptosis of cancer cells.
2021, 28(10):969-977.
Abstract:
Objective: To explore the effect of lycorine hydrochloride (LH) on the malignant biological behaviors of liver cancer HCCLM3 cells and its regulation on circASH2L/miR-124-3p axis. Methods: HCCLM3 cells treated with different concentrations of LH were divided into LH groups with different concentrations (LH-L group, LH-M group and LH-H group), Con group, si-NC group, si-circASH2L group, LH+pcDNA group, LH+pcDNA-circASH2L group. CCK-8 assay, plate clone formation test, flow cytometry, wound-healing assay, and Transwell assay were used to detect the proliferation, clone formation, apoptosis, migration and invasion of HCCLM3 cells. qPCR was used to detect the expression levels of circASH2L and miR-124-3p in HCCLM3 cells. Dual luciferase reporter gene assay was used to experiment detect the targeting relationship between circASH2L and miR-124-3p. WB was used to detect the expression of cleaved-caspase3, cleaved-caspase9, E-cadherin, and N-cadherin.Results: Compared with the Con group, the cell proliferation inhibition rates of LH groups with different concentrations were increased (P<0.05), the apoptosis rates and the expression levels of cleaved-caspase3, cleaved-caspase9 and E-cadherin were increased (all P<0.05), the expression levels of miR-124-3p were increased (P<0.05), the number of clone formation and invasive cells was decreased (all P<0.05), the wound-healing rates and the expression levels of N-cadherin were decreased (all P<0.05), and the expression levels of circASH2L were decreased (P<0.05), and the difference among different concentration groups was statistically significant(all P<0.05). CircASH2L could negatively regulate miR-124-3p. Compared with the si-NC group, the cell proliferation inhibition rates of the si-circASH2L group was increased (P<0.05), and the apoptosis rate and the expression levels of cleaved-caspase3, cleaved-caspase9 and E-cadherin were increased (all P<0.05), the wound-healing rate and the expression levels of N-cadherin were decreased (all P<0.05), and the number of clone formation and invasive cells was decreased (all P<0.05). Compared with the LH+pcDNA group, the expression level of miR-124-3p in the LH+ pcDNA-circASH2L group was decreased (P<0.05), the cell proliferation inhibition rate was decreased (P<0.05), the apoptosis rate and the expression levels of cleaved-caspase3, cleaved-caspase9 and E-cadherin were decreased (all P<0.05), the number of clone formation and invasive cells was increased (P<0.05), and the wound-healing rate and expression level of N-cadherin were increased (all P<0.05).
Objective: To explore the effect of lycorine hydrochloride (LH) on the malignant biological behaviors of liver cancer HCCLM3 cells and its regulation on circASH2L/miR-124-3p axis. Methods: HCCLM3 cells treated with different concentrations of LH were divided into LH groups with different concentrations (LH-L group, LH-M group and LH-H group), Con group, si-NC group, si-circASH2L group, LH+pcDNA group, LH+pcDNA-circASH2L group. CCK-8 assay, plate clone formation test, flow cytometry, wound-healing assay, and Transwell assay were used to detect the proliferation, clone formation, apoptosis, migration and invasion of HCCLM3 cells. qPCR was used to detect the expression levels of circASH2L and miR-124-3p in HCCLM3 cells. Dual luciferase reporter gene assay was used to experiment detect the targeting relationship between circASH2L and miR-124-3p. WB was used to detect the expression of cleaved-caspase3, cleaved-caspase9, E-cadherin, and N-cadherin.Results: Compared with the Con group, the cell proliferation inhibition rates of LH groups with different concentrations were increased (P<0.05), the apoptosis rates and the expression levels of cleaved-caspase3, cleaved-caspase9 and E-cadherin were increased (all P<0.05), the expression levels of miR-124-3p were increased (P<0.05), the number of clone formation and invasive cells was decreased (all P<0.05), the wound-healing rates and the expression levels of N-cadherin were decreased (all P<0.05), and the expression levels of circASH2L were decreased (P<0.05), and the difference among different concentration groups was statistically significant(all P<0.05). CircASH2L could negatively regulate miR-124-3p. Compared with the si-NC group, the cell proliferation inhibition rates of the si-circASH2L group was increased (P<0.05), and the apoptosis rate and the expression levels of cleaved-caspase3, cleaved-caspase9 and E-cadherin were increased (all P<0.05), the wound-healing rate and the expression levels of N-cadherin were decreased (all P<0.05), and the number of clone formation and invasive cells was decreased (all P<0.05). Compared with the LH+pcDNA group, the expression level of miR-124-3p in the LH+ pcDNA-circASH2L group was decreased (P<0.05), the cell proliferation inhibition rate was decreased (P<0.05), the apoptosis rate and the expression levels of cleaved-caspase3, cleaved-caspase9 and E-cadherin were decreased (all P<0.05), the number of clone formation and invasive cells was increased (P<0.05), and the wound-healing rate and expression level of N-cadherin were increased (all P<0.05).
ZHU Yonggang
,
TIAN Ziqiang
,
ZHAO Zhenxiang
,
WANG Mingbo
,
CAO Feng
,
WEN Shiwang
,
LI Zhenhua
,
SHAN Baoen
2021, 28(10):978-984.
Abstract:
Objective: To investigate the expression of miR-627-3p in esophageal squamous cell carcinoma (ESCC) tissues and its effect on biological behaviours of ESCC cells. Methods: From January 2015 to October 2015, 86 ESCC tissue specimens and 20 cooresponding adjacent normal tissue specimens from Thoracic Surgery Department of the Fourth Hospital of Hebei Medical University were collected. qPCR was used to detect the expression level of miR-627-3p in 86 cases of ESCC tissues and 20 cases of adjacent normal tissues. The relationship between miR-627-3p expression and clinicopathological parameters as well as prognosis was analyzed. The Kaplan-Meier plotter online database was used to further analyze the relationship between hsa-miR-627 expression and the prognosis of ESCC patients. qPCR was used to detect the expression of miR-627-3p in the four ESCC cells. miR-627-3p mimic was transfected into low-expressed cells, and miR-627-3p inhibitor was transfected into high-expressed cells. The cell proliferation was detected by CCK-8 assay, and the cell migration and invasion were detected by Transwell assay, respectively. KEGG analysis was used to explore the possible signal pathways mediated by miR-627-3p, and qPCR was used to verify the effect of miR-627-3p on the expression of key genes in the siganl pathway. Results: The expression of miR-627-3p in ESCC tissues was significantly lower than that in adjacent normal tissues. The expression of miR-627-3p was negatively correlated with lymph node metastasis and clinical stage of ESCC patients (all P<0.05). The five-year survival rate of ESCC patients with high miR-627-3p expression was significantly higher than that of the patients with low miR-627-3p expression (P<0.05). The expression of miR-627-3p was lowest in KYSE170 cells, and highest in KYSE30 cells. After transfection of miR-627-3p mimic into KYSE170 cells, the cell proliferation ability was not changed (P>0.05), but cell migration (P<0.05) and invasion (P<0.05) abilities were significantly decreased. After transfection of miR-627-3p inhibitor into KYSE30 cells, the cell proliferation ability was not changed (P>0.05), but cell migration (P<0.05) and invasion (P<0.05)abilities were significantly increased. KEGG analysis showed that miR-627-3p mediated multiple tumor-related signal pathways.Conclusion: The expression of miR-627-3p in ESCC tissues is significantly lower than in adjacent normal tissues, and its low expression is related to the poor prognosis of ESCC patients. miR-627-3p inhibits the migration and invasion of ESCC cells, which might play a biological function by interfering with multiple tumor-related signal pathways.
Objective: To investigate the expression of miR-627-3p in esophageal squamous cell carcinoma (ESCC) tissues and its effect on biological behaviours of ESCC cells. Methods: From January 2015 to October 2015, 86 ESCC tissue specimens and 20 cooresponding adjacent normal tissue specimens from Thoracic Surgery Department of the Fourth Hospital of Hebei Medical University were collected. qPCR was used to detect the expression level of miR-627-3p in 86 cases of ESCC tissues and 20 cases of adjacent normal tissues. The relationship between miR-627-3p expression and clinicopathological parameters as well as prognosis was analyzed. The Kaplan-Meier plotter online database was used to further analyze the relationship between hsa-miR-627 expression and the prognosis of ESCC patients. qPCR was used to detect the expression of miR-627-3p in the four ESCC cells. miR-627-3p mimic was transfected into low-expressed cells, and miR-627-3p inhibitor was transfected into high-expressed cells. The cell proliferation was detected by CCK-8 assay, and the cell migration and invasion were detected by Transwell assay, respectively. KEGG analysis was used to explore the possible signal pathways mediated by miR-627-3p, and qPCR was used to verify the effect of miR-627-3p on the expression of key genes in the siganl pathway. Results: The expression of miR-627-3p in ESCC tissues was significantly lower than that in adjacent normal tissues. The expression of miR-627-3p was negatively correlated with lymph node metastasis and clinical stage of ESCC patients (all P<0.05). The five-year survival rate of ESCC patients with high miR-627-3p expression was significantly higher than that of the patients with low miR-627-3p expression (P<0.05). The expression of miR-627-3p was lowest in KYSE170 cells, and highest in KYSE30 cells. After transfection of miR-627-3p mimic into KYSE170 cells, the cell proliferation ability was not changed (P>0.05), but cell migration (P<0.05) and invasion (P<0.05) abilities were significantly decreased. After transfection of miR-627-3p inhibitor into KYSE30 cells, the cell proliferation ability was not changed (P>0.05), but cell migration (P<0.05) and invasion (P<0.05)abilities were significantly increased. KEGG analysis showed that miR-627-3p mediated multiple tumor-related signal pathways.Conclusion: The expression of miR-627-3p in ESCC tissues is significantly lower than in adjacent normal tissues, and its low expression is related to the poor prognosis of ESCC patients. miR-627-3p inhibits the migration and invasion of ESCC cells, which might play a biological function by interfering with multiple tumor-related signal pathways.
2021, 28(10):985-989.
Abstract:
Objective: To explore the expression of F-box only protein 2 (FBXO2) gene in human gastric cancer cell lines, and its effect on the proliferation, migration, invasion and epithelial-mesenchymal transition (EMT) of gastric cancer cells.Methods: Gastric cancer cell lines (MGC-803, AGS, SGC-7901, MKN-28) and normal gastric epithelial cell line GES-1 were selected for this study; and Real-time quantitative PCR (qPCR) was used to detected the expression level of FBXO2 mRNA in the five cell lines. Designed a specific siRNA to inhibit the expression of FBXO2 and transiently transfected into MGC-803 call, and set up a negative control group for transfection of siRNA nonsense sequences. qPCR was used to detect the expression level of FBXO2 mRNA in MGC-803 cells 48 h after transfection; MTT, wound scratch, and Transwell assays were used to detect the effects of FBXO2 down-regulation on cell proliferation, migration and invasion, and WB was used to detect the expression of EMT-related proteins E-cadherin, N-cadherin and vimentin in cells.Results: The expression level of FBXO2 mRNA in the four gastric cancer cell lines was significantly higher than that in gastric epithelial cell line GES-1 (P<0.05 or P<0.01). Compared with the negative control group, FBXO2 mRNA expression in the siRNA-FBXO2 group was decreased (P<0.01), and the proliferation, migration and invasion of the cells were significantly inhibited (P<0.05 or P<0.01), and E-cadherin protein expression was significantly increased (P<0.01), while N-cadherin and vimentin protein expression were significantly decreased (all P<0.01). Conclusion: The low expression of FBXO2 can efficiently inhibited the proliferation, migration and invasion of gastric cancer cells, which might be related with EMT.
Objective: To explore the expression of F-box only protein 2 (FBXO2) gene in human gastric cancer cell lines, and its effect on the proliferation, migration, invasion and epithelial-mesenchymal transition (EMT) of gastric cancer cells.Methods: Gastric cancer cell lines (MGC-803, AGS, SGC-7901, MKN-28) and normal gastric epithelial cell line GES-1 were selected for this study; and Real-time quantitative PCR (qPCR) was used to detected the expression level of FBXO2 mRNA in the five cell lines. Designed a specific siRNA to inhibit the expression of FBXO2 and transiently transfected into MGC-803 call, and set up a negative control group for transfection of siRNA nonsense sequences. qPCR was used to detect the expression level of FBXO2 mRNA in MGC-803 cells 48 h after transfection; MTT, wound scratch, and Transwell assays were used to detect the effects of FBXO2 down-regulation on cell proliferation, migration and invasion, and WB was used to detect the expression of EMT-related proteins E-cadherin, N-cadherin and vimentin in cells.Results: The expression level of FBXO2 mRNA in the four gastric cancer cell lines was significantly higher than that in gastric epithelial cell line GES-1 (P<0.05 or P<0.01). Compared with the negative control group, FBXO2 mRNA expression in the siRNA-FBXO2 group was decreased (P<0.01), and the proliferation, migration and invasion of the cells were significantly inhibited (P<0.05 or P<0.01), and E-cadherin protein expression was significantly increased (P<0.01), while N-cadherin and vimentin protein expression were significantly decreased (all P<0.01). Conclusion: The low expression of FBXO2 can efficiently inhibited the proliferation, migration and invasion of gastric cancer cells, which might be related with EMT.
2021, 28(10):990-997.
Abstract:
Objective: To search for potential pathogenic genes, disease diagnosis and prognosis related factors in multiple myeloma (MM) microarray datasets and clarify their mechanism. Methods: GSE13591 and Zhan Myeloma datasets were obtained, and the gene differential expression was analyzed by R language. Meta-analysis was performed based on merged datasets, and the TOP10 genes in median rank of P-value were screened. The mRNA expression of TOP10 genes in tumor cell lines were analyzed using CCLE database,and proteasome pathway related gene 8 (DCAF8) was screened. The mutation frequency of DCAF8 gene in each tumor cell lines was analyzed in cBioPortal database. Furthermore, the expression of DCAF8 protein in MM cell lines were detected by using WB. SPSS and Graph Pad software were employed to analyze the correlation between DCAF8 expression and clinical characteristics of MM patients. In addition, the receiver operating characteristic curve (ROC curve) and survival analysis were conducted according to the expression level of DCAF8. Finally, R clusterProfiler package and Metascape database were used to perform GO and KEGG functional enrichment analyses of DCAF8 interacting protein gene and co-expressed genes. Results: A total of 477 high expression genes were obtained from the intersection of the up-regulated genes of GSE13591 and Zhan Myeloma datasets. Meta-analysis of the above genes in the merged dataset was performed, the TOP10 genes in median P-value rank were HGF, AKAP1, TCTA, NEB, DCAF8, COPS6,MAP3K5, PON2, BAG1 and CD59. The analysis of CCLE database showed that the average expression level of DCAF8 was the highest in MM cell lines. The mutation frequency of DCAF8 gene in plasma cell myeloma ranked first among all tumor cell lines in cBioPortal database. The result of WB showed that the level of DCAF8 protein was increased significantly in a variety of MM cell lines compared with 293T control. The results showed that the expression of DCAF8 was positively correlated with the tumor load of MM patients, and the expression of DCAF8 in positive group of 1q21 amplification was significantly higher than that in negative group. The results of ROC curve based on GSE13591 and GSE16558 manifested that the expression level of DCAF8 had a good diagnostic value for MM (P<0.01). The survival analysis related to DCAF8 expression was conducted using the GSE2658, GSE4452 and GSE9782 datasets, and it showed that the level of DCAF8 expression was negatively correlated with the overall survival (OS) rate of MM patients. Protein interaction network showed that DCAF8 could interact with XPO1 protein directly. The GO and KEGG functional enrichment analysis showed that DCAF8 was related to proteasome function, spliceosome activity, histone acetylase activity, RNA transport, etc. Conclusion: DCAF8 is highly expressed in MM significantly, and its expression level can distinguish MM patients from normal people. Its expression is negatively correlated with the overall survival time of MM patients. Remarkably, DCAF8 is related to 1q21 amplification and could interact with XPO1 protein directly. Therefore, DCAF8 maybe a new biomarker of MM.
Objective: To search for potential pathogenic genes, disease diagnosis and prognosis related factors in multiple myeloma (MM) microarray datasets and clarify their mechanism. Methods: GSE13591 and Zhan Myeloma datasets were obtained, and the gene differential expression was analyzed by R language. Meta-analysis was performed based on merged datasets, and the TOP10 genes in median rank of P-value were screened. The mRNA expression of TOP10 genes in tumor cell lines were analyzed using CCLE database,and proteasome pathway related gene 8 (DCAF8) was screened. The mutation frequency of DCAF8 gene in each tumor cell lines was analyzed in cBioPortal database. Furthermore, the expression of DCAF8 protein in MM cell lines were detected by using WB. SPSS and Graph Pad software were employed to analyze the correlation between DCAF8 expression and clinical characteristics of MM patients. In addition, the receiver operating characteristic curve (ROC curve) and survival analysis were conducted according to the expression level of DCAF8. Finally, R clusterProfiler package and Metascape database were used to perform GO and KEGG functional enrichment analyses of DCAF8 interacting protein gene and co-expressed genes. Results: A total of 477 high expression genes were obtained from the intersection of the up-regulated genes of GSE13591 and Zhan Myeloma datasets. Meta-analysis of the above genes in the merged dataset was performed, the TOP10 genes in median P-value rank were HGF, AKAP1, TCTA, NEB, DCAF8, COPS6,MAP3K5, PON2, BAG1 and CD59. The analysis of CCLE database showed that the average expression level of DCAF8 was the highest in MM cell lines. The mutation frequency of DCAF8 gene in plasma cell myeloma ranked first among all tumor cell lines in cBioPortal database. The result of WB showed that the level of DCAF8 protein was increased significantly in a variety of MM cell lines compared with 293T control. The results showed that the expression of DCAF8 was positively correlated with the tumor load of MM patients, and the expression of DCAF8 in positive group of 1q21 amplification was significantly higher than that in negative group. The results of ROC curve based on GSE13591 and GSE16558 manifested that the expression level of DCAF8 had a good diagnostic value for MM (P<0.01). The survival analysis related to DCAF8 expression was conducted using the GSE2658, GSE4452 and GSE9782 datasets, and it showed that the level of DCAF8 expression was negatively correlated with the overall survival (OS) rate of MM patients. Protein interaction network showed that DCAF8 could interact with XPO1 protein directly. The GO and KEGG functional enrichment analysis showed that DCAF8 was related to proteasome function, spliceosome activity, histone acetylase activity, RNA transport, etc. Conclusion: DCAF8 is highly expressed in MM significantly, and its expression level can distinguish MM patients from normal people. Its expression is negatively correlated with the overall survival time of MM patients. Remarkably, DCAF8 is related to 1q21 amplification and could interact with XPO1 protein directly. Therefore, DCAF8 maybe a new biomarker of MM.
2021, 28(10):998-1004.
Abstract:
Objective: To explore the expression of transmembrane protein 125(TMEM125) in lung adenocarcinoma (LUAD) tissues and A549 cells, and the molecular mechanism that affects the proliferation and invasion of A549 cells. Methods: Collected and downloaded the lung adenocarcinoma clinical information and gene expression profile data from The Cancer Genome Atlas (TCGA) database, and analyzed the correlation between the expression of TMEM125 in lung adenocarcinoma and the overall survival of patients.Constructed TMEM125 overexpressing A549 cell line, detected the effect of TMEM125 overexpression on the proliferation and migration of A549 cells by CCK-8 assay and Wound healing assay, and detected the effect of TMEM125 overexpression on the cell cycle and apoptosis of A549 cells by using flow cytometry. WB was used to detect the effect of TMEM125 overexpression on downstream NF- κB signaling pathways and apoptotic proteins. Co-immunoprecipitation (Co-IP) was used to detect the interaction between TMEM125 and NF-κB inhibitor interacting Ras-like 2 (NKIRAS2). TMEM125 overexpressing cells was treated with TNFα（10 ng/ml）and then CCK-8 assay, flow cytometry and WB were used to detect its effects on cell proliferation, apoptosis and NF- κB signaling pathway proteins. A549 cells were treated with demethylation reagent decitabine, and the expression of TMEM125 gene and protein was detected by qPCR and WB. Results: The expression level of TMEM125 mRNA in LUAD tissue was significantly lower than that in normal tissues (P<0.001), the promoter methylation level was significantly higher than that in normal tissues (P<0.001), and the overall survival of patients with low and medium expression was significantly lower than that of patients with high expression (P<0.001).Overexpression of TMEM125 inhibited the proliferation and migration of A549 cells (P<0.01), increased cell G2/M phase and promoted cell apoptosis (P<0.01). Overexpression of TMEM125 could interact with NKIRAS2 and inhibit the activity of NF- ΚB (P<0.01).Treatment of A549 cells with decitabine could promote the expression of TMEM125 and inhibit cell proliferation (P<0.01).Conclusion:Promoter hypermethylation inhibits TMEM125 gene expression, leading to a decline in its function of inhibiting NF-κB activity and inhibiting cell proliferation and therefore reduce the sensitivity to decitabine, and resulting in the decreased sensitivity to decitabine.
Objective: To explore the expression of transmembrane protein 125(TMEM125) in lung adenocarcinoma (LUAD) tissues and A549 cells, and the molecular mechanism that affects the proliferation and invasion of A549 cells. Methods: Collected and downloaded the lung adenocarcinoma clinical information and gene expression profile data from The Cancer Genome Atlas (TCGA) database, and analyzed the correlation between the expression of TMEM125 in lung adenocarcinoma and the overall survival of patients.Constructed TMEM125 overexpressing A549 cell line, detected the effect of TMEM125 overexpression on the proliferation and migration of A549 cells by CCK-8 assay and Wound healing assay, and detected the effect of TMEM125 overexpression on the cell cycle and apoptosis of A549 cells by using flow cytometry. WB was used to detect the effect of TMEM125 overexpression on downstream NF- κB signaling pathways and apoptotic proteins. Co-immunoprecipitation (Co-IP) was used to detect the interaction between TMEM125 and NF-κB inhibitor interacting Ras-like 2 (NKIRAS2). TMEM125 overexpressing cells was treated with TNFα（10 ng/ml）and then CCK-8 assay, flow cytometry and WB were used to detect its effects on cell proliferation, apoptosis and NF- κB signaling pathway proteins. A549 cells were treated with demethylation reagent decitabine, and the expression of TMEM125 gene and protein was detected by qPCR and WB. Results: The expression level of TMEM125 mRNA in LUAD tissue was significantly lower than that in normal tissues (P<0.001), the promoter methylation level was significantly higher than that in normal tissues (P<0.001), and the overall survival of patients with low and medium expression was significantly lower than that of patients with high expression (P<0.001).Overexpression of TMEM125 inhibited the proliferation and migration of A549 cells (P<0.01), increased cell G2/M phase and promoted cell apoptosis (P<0.01). Overexpression of TMEM125 could interact with NKIRAS2 and inhibit the activity of NF- ΚB (P<0.01).Treatment of A549 cells with decitabine could promote the expression of TMEM125 and inhibit cell proliferation (P<0.01).Conclusion:Promoter hypermethylation inhibits TMEM125 gene expression, leading to a decline in its function of inhibiting NF-κB activity and inhibiting cell proliferation and therefore reduce the sensitivity to decitabine, and resulting in the decreased sensitivity to decitabine.
2021, 28(10):1005-1014.
Abstract:
Objective: To explore the mechanism of linc00941 adsorbing miR-203 as ceRNA and up-regulating the expression of CC chemokine ligand 2 (CCL2) in esophageal squamous cell carcinoma (ESCC). Methods: The cancer tissues and adjacent tissues of 58 ESCC patients in the First Affiliated Hospital of Nanyang Medical College were selected, including 33 male patients aged (49.3±18.6) years and 25 female patients aged (44.6±20.7) years. The differential expression of linc00941, miR-203 and CCL2 in ESCC tissue, four human ESCC cell lines (EC9706, KYSE30, ECA109 and TE1) and human normal esophageal epithelial cell line HET-1A were detected by qRT-PCR. linc00941-wt, linc00941-mut, CCL2-wt and CCL2-mut plasmids were constructed and co transfected into 293T cells with miR-203 NC or miR-203 mimic, respectively. Dual luciferase reporter gene assay was implemented to verify the interaction between linc00941, miR-203 and CCL2. In addition, CCK-8 and Transwell experiments were used to detect the proliferation and invasion of cells. The lactic acid (LA) content was measured to evaluate the glycolysis ability of the cells. The cell apoptosis was detected by flow cytometry. Glycolysis inhibitor 2-DG and linc00941 were used to intervene ESCC cells to further observe the regulatory effect of linc00941 on ESCC cells. Results: The expressions of linc00941 and CCL2 were up-regulated while the expression of miR-203 was down-regulated in ESCC tissues and cell lines (all P<0.05). The interactions of linc00941 with miR-203 and miR-203 with CCL2 were both confirmed in ECA109 cells. Knockdown of linc00941 could inhibit the proliferation, invasion, glycolysis of ECA109 cells and induce their apoptosis, which was partly reversed by miR-203 inhibitor (all P<0.05). At the same time, CCL2 overexpression could partly reverse the effects of knockdown of linc00941 on the proliferation, invasion, glycolysis and apoptosis of ECA109 cells (all P<0.05). Conclusion: linc00941 can increase the expression of CCL2 via absorbing miR-203, subsequently promote the proliferation,invasion, glycolysis of ESCC cells and induce their apoptosis. The effects of linc00941 on the proliferation, invasion and apoptosis of ESCC cells may be realized by regulating glycolysis.
Objective: To explore the mechanism of linc00941 adsorbing miR-203 as ceRNA and up-regulating the expression of CC chemokine ligand 2 (CCL2) in esophageal squamous cell carcinoma (ESCC). Methods: The cancer tissues and adjacent tissues of 58 ESCC patients in the First Affiliated Hospital of Nanyang Medical College were selected, including 33 male patients aged (49.3±18.6) years and 25 female patients aged (44.6±20.7) years. The differential expression of linc00941, miR-203 and CCL2 in ESCC tissue, four human ESCC cell lines (EC9706, KYSE30, ECA109 and TE1) and human normal esophageal epithelial cell line HET-1A were detected by qRT-PCR. linc00941-wt, linc00941-mut, CCL2-wt and CCL2-mut plasmids were constructed and co transfected into 293T cells with miR-203 NC or miR-203 mimic, respectively. Dual luciferase reporter gene assay was implemented to verify the interaction between linc00941, miR-203 and CCL2. In addition, CCK-8 and Transwell experiments were used to detect the proliferation and invasion of cells. The lactic acid (LA) content was measured to evaluate the glycolysis ability of the cells. The cell apoptosis was detected by flow cytometry. Glycolysis inhibitor 2-DG and linc00941 were used to intervene ESCC cells to further observe the regulatory effect of linc00941 on ESCC cells. Results: The expressions of linc00941 and CCL2 were up-regulated while the expression of miR-203 was down-regulated in ESCC tissues and cell lines (all P<0.05). The interactions of linc00941 with miR-203 and miR-203 with CCL2 were both confirmed in ECA109 cells. Knockdown of linc00941 could inhibit the proliferation, invasion, glycolysis of ECA109 cells and induce their apoptosis, which was partly reversed by miR-203 inhibitor (all P<0.05). At the same time, CCL2 overexpression could partly reverse the effects of knockdown of linc00941 on the proliferation, invasion, glycolysis and apoptosis of ECA109 cells (all P<0.05). Conclusion: linc00941 can increase the expression of CCL2 via absorbing miR-203, subsequently promote the proliferation,invasion, glycolysis of ESCC cells and induce their apoptosis. The effects of linc00941 on the proliferation, invasion and apoptosis of ESCC cells may be realized by regulating glycolysis.
8 Down-regulation of miR-183 targeted tumor suppressor CPEB1 enhances radiosensitivity of glioma cells
YANG Sen
,
DONG Lixin
,
ZHANG Yanqiu
,
FU Baohong
,
CAO Liyan
,
MAO Yu
,
HANG Qinghuai
,
WANG Guangxia
,
FU Zhanzhao
,
WU Lei
,
WANG Dong
2021, 28(10):1015-1022.
Abstract:
Objective: To investigate the effect of miR-183 on radiosensitivity of glioma cells.Methods: From October 2020 to June 2021, tissue samples of 40 cases of brain glioma from the First Hospital of Qinhuangdao were collected. T98G cells were irradiated with gradient dose X-ray (0, 2, 4, 6 Gy). The expression of miR-183 and CPEB1 in glioma tissues, T98G cells and X-ray irradiated T98G cells were detected by qPCR. After transfection of T98G cells with miR-183 inhibitor, the expression of miR-183 was down-regulated. The proliferation ability, apoptosis rate, BAX and Bcl2 protein expression of T98G cells were detected by CCK-8 assay, flow cytometry and WB, respectively, after 6 Gy X-ray vertical irradiation. The targeting relationship between miR-183 and CPEB1 was detected by Targetscan software prediction and dual luciferase reporter gene assay. After down regulating the expression of CPEB1, after 6 Gy X-ray irradiation, the proliferation ability, apoptosis rate, BAX and Bcl2 protein expression of T98G cells were detected by CCK-8 assay, flow cytometry and WB, respectively. T98G cells were transfected with pcDNA-CPEB1 or CPEB1 siRNA plasmid, and after CPEB1 was down-regulated or overexpressed, respectively, the effect of miR-183 on radiosensitivity of T98G cells through CPEB1 was detected.Results: The expression of miR-183 in glioma tissues and cells increased significantly, and the expression of CPEB1 mRNA decreased significantly. The expression of miR-183 in T98G cells decreased with the increase of X-ray radiation dose (P<0.05), and the expression of CPEB1 increased with the increase of X-ray radiation dose (P<0.05). After 6 Gy X-ray irradiation of T98G cells, down-regulation of miR-183 could reduce cell proliferation and increase apoptosis rate, while overexpression of miR-183 had the opposite effect (P<0.05).MiR-183 targeted mRNA CPEB1 and negatively regulated CPEB1 expression. After down-regulation of CPEB1, 6 Gy X-ray irradiation could significantly improve the proliferation of T98G cells (P<0.05) and reduce the apoptosis rate (P<0.05). MiR-183 could reverse the promoting effect of CPEB1 overexpression on radiosensitivity of T98G cells (P<0.05). Conclusion: Down-regulation of miR-183 expression can negatively regulate CPEB1, which might enhance the radiosensitivity of glioma cells.
Objective: To investigate the effect of miR-183 on radiosensitivity of glioma cells.Methods: From October 2020 to June 2021, tissue samples of 40 cases of brain glioma from the First Hospital of Qinhuangdao were collected. T98G cells were irradiated with gradient dose X-ray (0, 2, 4, 6 Gy). The expression of miR-183 and CPEB1 in glioma tissues, T98G cells and X-ray irradiated T98G cells were detected by qPCR. After transfection of T98G cells with miR-183 inhibitor, the expression of miR-183 was down-regulated. The proliferation ability, apoptosis rate, BAX and Bcl2 protein expression of T98G cells were detected by CCK-8 assay, flow cytometry and WB, respectively, after 6 Gy X-ray vertical irradiation. The targeting relationship between miR-183 and CPEB1 was detected by Targetscan software prediction and dual luciferase reporter gene assay. After down regulating the expression of CPEB1, after 6 Gy X-ray irradiation, the proliferation ability, apoptosis rate, BAX and Bcl2 protein expression of T98G cells were detected by CCK-8 assay, flow cytometry and WB, respectively. T98G cells were transfected with pcDNA-CPEB1 or CPEB1 siRNA plasmid, and after CPEB1 was down-regulated or overexpressed, respectively, the effect of miR-183 on radiosensitivity of T98G cells through CPEB1 was detected.Results: The expression of miR-183 in glioma tissues and cells increased significantly, and the expression of CPEB1 mRNA decreased significantly. The expression of miR-183 in T98G cells decreased with the increase of X-ray radiation dose (P<0.05), and the expression of CPEB1 increased with the increase of X-ray radiation dose (P<0.05). After 6 Gy X-ray irradiation of T98G cells, down-regulation of miR-183 could reduce cell proliferation and increase apoptosis rate, while overexpression of miR-183 had the opposite effect (P<0.05).MiR-183 targeted mRNA CPEB1 and negatively regulated CPEB1 expression. After down-regulation of CPEB1, 6 Gy X-ray irradiation could significantly improve the proliferation of T98G cells (P<0.05) and reduce the apoptosis rate (P<0.05). MiR-183 could reverse the promoting effect of CPEB1 overexpression on radiosensitivity of T98G cells (P<0.05). Conclusion: Down-regulation of miR-183 expression can negatively regulate CPEB1, which might enhance the radiosensitivity of glioma cells.
2021, 28(10):1023-1028.
Abstract:
调节性 B 细胞（Breg）是近年来新发现的 B 细胞功能型亚群，其通过分泌白细胞介素-10（IL-10）、白细胞介素-35 （IL-35）和转化生长因子-β（TGF-β）等负向调节因子抑制机体免疫应答，在肿瘤免疫逃逸中发挥重要作用，影响肿瘤的发展、侵袭 和转移。在肝癌、胃癌、乳腺癌、黑色素瘤、卵巢癌、肺癌等多种常见癌中均发现存在具有促癌效应的Breg。Breg和肿瘤的临床分 期、治疗、预后存在一定的相关性。靶向Breg可能成为未来肿瘤免疫治疗的一个新方向。
调节性 B 细胞（Breg）是近年来新发现的 B 细胞功能型亚群，其通过分泌白细胞介素-10（IL-10）、白细胞介素-35 （IL-35）和转化生长因子-β（TGF-β）等负向调节因子抑制机体免疫应答，在肿瘤免疫逃逸中发挥重要作用，影响肿瘤的发展、侵袭 和转移。在肝癌、胃癌、乳腺癌、黑色素瘤、卵巢癌、肺癌等多种常见癌中均发现存在具有促癌效应的Breg。Breg和肿瘤的临床分 期、治疗、预后存在一定的相关性。靶向Breg可能成为未来肿瘤免疫治疗的一个新方向。
2021, 28(10):1029-1036.
Abstract:
PD-1/PD-L1 免疫检查点抑制剂（ICI）在单药或联合治疗肝细胞癌（HCC）的临床试验显示出较好的疗效和安全 性，为其临床应用提供有利的科学依据。单药治疗 HCC 的 PD-1/PD-L1 ICI 包括纳武利尤单抗（nivolumab）、帕博利珠单抗 （pembrolizumab）和卡瑞利珠单抗（camrelizumab），主要用于肝癌的二线治疗。虽部分 HCC 患者可以对 PD-1/PD-L1 ICI 单药 治疗产生持久的反应，但总体受益的患者仍然较少。PD-1/D-L1 ICI 联合治疗显示出更好的免疫应答和控制。目前，常用的 联合治疗手段包括与其他类 ICI、靶向药物、放疗、化疗以及介入治疗等联合。与双 ICI 或 ICI 和化疗联合治疗相比，酪氨酸激 酶抑制剂（TKI）和 PD-1/PD-L1 ICI 联合治疗显示出更好的疗效。在安全性方面，联合治疗的不良反应发生率发也高于单药 治疗，且更易出现严重或致死性的不良反应。PD-1/PD-L1 ICI 单药和联合治疗 HBV/HCV 相关 HCC 患者的安全性和有效性 正在研究中，同时接受抗病毒药物治疗患者表现出稳定的治疗应答，但 HBV/HCV 病毒重新激活相关的安全性问题，尚未得 出一致结论。
PD-1/PD-L1 免疫检查点抑制剂（ICI）在单药或联合治疗肝细胞癌（HCC）的临床试验显示出较好的疗效和安全 性，为其临床应用提供有利的科学依据。单药治疗 HCC 的 PD-1/PD-L1 ICI 包括纳武利尤单抗（nivolumab）、帕博利珠单抗 （pembrolizumab）和卡瑞利珠单抗（camrelizumab），主要用于肝癌的二线治疗。虽部分 HCC 患者可以对 PD-1/PD-L1 ICI 单药 治疗产生持久的反应，但总体受益的患者仍然较少。PD-1/D-L1 ICI 联合治疗显示出更好的免疫应答和控制。目前，常用的 联合治疗手段包括与其他类 ICI、靶向药物、放疗、化疗以及介入治疗等联合。与双 ICI 或 ICI 和化疗联合治疗相比，酪氨酸激 酶抑制剂（TKI）和 PD-1/PD-L1 ICI 联合治疗显示出更好的疗效。在安全性方面，联合治疗的不良反应发生率发也高于单药 治疗，且更易出现严重或致死性的不良反应。PD-1/PD-L1 ICI 单药和联合治疗 HBV/HCV 相关 HCC 患者的安全性和有效性 正在研究中，同时接受抗病毒药物治疗患者表现出稳定的治疗应答，但 HBV/HCV 病毒重新激活相关的安全性问题，尚未得 出一致结论。
2021, 28(10):1037-1040.
Abstract:
结直肠癌（CRC）分子分型的方法与研究进展主要从基因组学、转录组学和蛋白质组学3个方面展开。依据基因突变 和基因组的细胞遗传学改变进行分型，可以将CRC分为错配修复缺陷导致微卫星不稳定的超突变肿瘤、具有DNA聚合酶epsilon 或Delta 1核酸外切酶结构域突变的超突变肿瘤、具有高频率DNA体细胞拷贝数改变的染色体不稳定性肿瘤等。基于肿瘤的上 皮间质转化、错配修复基因缺陷导致的微卫星不稳定性和细胞增殖相关的高突变频率，可分为MMR缺陷型上皮亚型（A型）、增 殖性上皮亚型（B型）和间充质亚型（C型）。根据基因表达谱的差异分型，可基于临床和组织病理学参数及基因特征、CRC基因表 达谱、全基因组mRNA表达亚型、对表皮生长因子受体靶向药物西妥昔单抗的治疗反应、共识分子等进行CRC亚型分类。基于 生物标志物研究的蛋白质组学分型，可分为亚型A—E等5种不同蛋白质组学亚型。分子分型有助于CRC个体化和精确化的治 疗策略。
结直肠癌（CRC）分子分型的方法与研究进展主要从基因组学、转录组学和蛋白质组学3个方面展开。依据基因突变 和基因组的细胞遗传学改变进行分型，可以将CRC分为错配修复缺陷导致微卫星不稳定的超突变肿瘤、具有DNA聚合酶epsilon 或Delta 1核酸外切酶结构域突变的超突变肿瘤、具有高频率DNA体细胞拷贝数改变的染色体不稳定性肿瘤等。基于肿瘤的上 皮间质转化、错配修复基因缺陷导致的微卫星不稳定性和细胞增殖相关的高突变频率，可分为MMR缺陷型上皮亚型（A型）、增 殖性上皮亚型（B型）和间充质亚型（C型）。根据基因表达谱的差异分型，可基于临床和组织病理学参数及基因特征、CRC基因表 达谱、全基因组mRNA表达亚型、对表皮生长因子受体靶向药物西妥昔单抗的治疗反应、共识分子等进行CRC亚型分类。基于 生物标志物研究的蛋白质组学分型，可分为亚型A—E等5种不同蛋白质组学亚型。分子分型有助于CRC个体化和精确化的治 疗策略。
2021, 28(10):1041-1048.
Abstract:
PD-1/PD-L1抑制剂在晚期胃癌（GC）及胃食管交界癌（GEJC）的临床试验中显示出较好的疗效及安全性，为其临床应 用提供了有力的科学依据。单药PD-1/PD-L1抑制剂治疗的疗效欠佳，仅作为后线治疗选择。PD-1/PD-L1抑制剂联合多种治疗 模式能够提高患者的疾病应答率和延长患者生存时间，成为目前GC及GEJC治疗的热点。本文基于最新临床试验结果，对晚期 GC及GEJC的免疫治疗疗效及预测分子进行综述。
PD-1/PD-L1抑制剂在晚期胃癌（GC）及胃食管交界癌（GEJC）的临床试验中显示出较好的疗效及安全性，为其临床应 用提供了有力的科学依据。单药PD-1/PD-L1抑制剂治疗的疗效欠佳，仅作为后线治疗选择。PD-1/PD-L1抑制剂联合多种治疗 模式能够提高患者的疾病应答率和延长患者生存时间，成为目前GC及GEJC治疗的热点。本文基于最新临床试验结果，对晚期 GC及GEJC的免疫治疗疗效及预测分子进行综述。
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Chinese Journal Of Cancer Biotheray ® 2025 All Rights Reserved
Supported by:Beijing E-Tiller Technology Development Co., Ltd.