COL5A2在胃癌组织中表达及其对胃癌细胞增殖、侵袭能力的影响

Expression of COL5A2 in gastric cancer tissues and its effect on proliferation and invasion ability of gastric cancer cells

目的:探讨Ⅴ型胶原蛋白α2(COL5A2)在胃癌组织中表达情况及其对胃癌细胞BGC823表型影响,并研究其相关影响机制。方法:收集手术切除的胃癌组织及相应癌旁正常组织各18例,通过实时荧光定量PCR(qRT-PCR)及Western blotting检测COL5A2表达差异。人胃癌细胞株BGC823转染siRNA,敲低COL5A2基因在细胞中表达量,通过qRT-PCR及Western blotting验证转染效率。通过克隆形成实验检测各组细胞增殖变化,Transwell检测细胞侵袭变化,Western blotting检测细胞中Wnt/β-catenin信号通路相关蛋白MMP9、Wnt、β-catenin表达。结果:与癌旁正常组织相比,胃癌组织中COL5A2蛋白表达增高(P<0.05)。沉默COL5A2表达后,BGC823细胞增殖及侵袭能力均降低(P<0.05),且MMP9、Wnt、β-catenin蛋白表达量均下调(P<0.05)。结论:COL5A2蛋白表达在胃癌组织中表达上调,并且可能通过Wnt/β-catenin信号通路影响胃癌细胞增殖及侵袭。

Objective:To investigate the expression of collagen type Vα2(COL5A2)in gastric cancer and its effect on the phenotype of BGC823 cells,and to study the related mechanism.Methods:A total of 18 cases of surgically removed gastric cancer tissues and corresponding adjacent normal tissues were collected.The expression difference of COL5A2 was detected by real-time quantitative fluorescent PCR(qRT-PCR)and Western blotting.Human gastric cancer cell line BGC823 was transfected with siRNA,and the expression of COL5A2 gene was knocked down in the cells.The transfection efficiency was verified by qRT-PCR and Western blotting.Clonal formation assay was used to detect cell proliferation in each group,and Transwell assay was used to detect cell invasion.The expression levels of Wnt/β-catenin signaling pathway-related proteins(MMP9,Wnt,β-catenin)were detected by Western blotting.Results:Compared with adjacent normal tissues,the expression of COL5A2 protein in gastric cancer tissues was increased(P<0.05).After the expression of COL5A2 was silenced,the proliferation and invasion ability of BGC823 cells were decreased(P<0.05),and the expression levels of MMP9,Wnt andβ-catenin were down-regulated(P<0.05).Conclusions:The expression of COL5A2 protein is up-regulated in gastric cancer tissues,and may affect the proliferation and invasion of gastric cancer cells through Wnt/β-catenin signaling pathway.