中国临床药理学与治疗学2017，Vol.22Issue(4)：401-405,5.
胃泌素调控ERK信号通路在促进大肠癌CACO2细胞增殖中的作用
Effects of gastrin on ERK signaling pathway in the proliferation of colorectal cancer cell line CACO2
摘要
Abstract
AIM:To investigate the effects and mechanisms of exrtacellular-signal regulated protein kinase in gastrin-induced cell proliferation of colorectal cancer cells by using RNA interference technology.METHODS:CCK-BR positive cell strain CACO2 was constructed lentivirus infection.Detection of gastrin receptor (CCK-BR) expression in colorectal cancer cell line CACO2 by qRT-PCR.RNA interference was used to silence the ERK1/2 gene,and the expression of total ERK protein and its phosphorylation level were analyzed.The experiment was divided into four groups:The control group,negative interference group,gastrin group and plasmid interference group.Changes of proliferation index of CACO2 cells were detected by cell cytometry.The protein of ERK1/2 and phosphorylation levels were detected by Western blot.RESULTS:After chronic viral infection,the target strip membrane protein and mRNA expression of CCK-BR were detected in the CACO2 cells.The amplified product was 185 bp.The interference plasmid was selected,the ratio of plasmid and liposome was 1 ∶ 2,the transfection efficiency was 40％-50％,and the expression level of ERK protein in the transfected plasmid was decreased.From the protein level,the constructed plasmid was observed effective.The proliferation index of the gastrin group was significantly higher than that of the plasmid interference group,the negative interference group and the blank control group,the difference was statistically significant (P ＜ 0.01).The expression of ERK1/2 protein and phosphorylation level in the four groups were significantly different (P ＜ 0.01).The protein expression and phosphorylation level of ERK1/2 in gastrin group were significantly higher than those in the plasmid interference group,the negative interference group and the blank control group,the difference was statistically significant (P ＜ 0.01).CONCLUSION:Gastrin can promote the proliferation of CACO2 cells in vitro by ERK signaling pathway.RNA interference can effectively block the phosphorylation of ERK protein in ERK signaling pathway,which is expected to provide a new approach for the treatment of colorectal cancer.关键词
胃泌素/ERK1/2/大肠癌/RNA干扰/慢病毒感染Key words
gastrin/ERK1/2/colorectal cancer/RNAi/lentivirus infection分类
医药卫生引用本文复制引用
茆家定,胡迪,吴佩..胃泌素调控ERK信号通路在促进大肠癌CACO2细胞增殖中的作用[J].中国临床药理学与治疗学,2017,22(4):401-405,5.基金项目
安徽省自然科学基金资助项目(1408085MH148) （1408085MH148）
皖南医学院弋矾山医院人才引进基金(YR201406) （YR201406）
