经典瞬时受体电位通道 1在胃癌组织中的表达及其生物学意义

目的 研究胃癌组织中经典瞬时受体电位通道1(transient receptor potential canonical,TRPC1)的表达及TRPC1沉默对胃癌细胞的生物学效应.方法 收集咸宁市中心医院2015年1月至2018年4月93例手术切除的胃癌和癌旁组织标本,免疫组化检测上述标本中TRPC1的表达,分析其与病人临床病理特征的关系.利用TRPC1小分子干扰RNA(siRNA-TRPC1)转染胃癌MGC-803细胞,逆转录聚合酶链反应(RT-PCR)和蛋白质印迹法(Western blotting)验证转染效率.MTT法评估siRNA-TRPC1对MGC-803细胞增殖能力的影响.相关试剂盒检测丙二醛、活性氧和超氧化物歧化酶(SOD)水平.流式细胞术检测细胞凋亡水平.结果 与癌旁组织比较,TRPC1在胃癌组织中的表达显著上调(阳性表达率分别为79.6％比20.4％),且TRPC1的高表达与肿瘤的TNM分期(P=0.007)、分化程度(P=0.022)和淋巴结转移(P=0.010)相关;与对照组比较,转染组TRPC1 mRNA和蛋白质水平均显著下降(P<0.05);与对照组比较,siRNA-TRPC1转染组细胞的增殖活性显著下降(P<0.05);与对照组比较,siRNA-TRPC1转染组细胞丙二醛和活性氧水平升高,SOD活性降低(P<0.05);与对照组比较,siRNA-TRPC1转染组细胞凋亡水平明显升高(P<0.05).结论 TRPC1在胃癌中高表达,siRNA沉默TRPC1可抑制胃癌肿瘤细胞的增殖,上调细胞的氧化应激水平,促进肿瘤细胞的凋亡.

The expression and biological significance of TRPC1 in gastric carcinoma tissue

Objective To investigate the expression and biological functions of transient receptor potential canonical 1 (TRPC1) in gastric carcinoma.Methods Ninety-three cases of surgically removed gastric carcinoma and paracamcinoma tissues in Xianning Central Hospital January 2015 to April 2018 were collected. The expression of TRPC1 in the above samples was detected by immunohistochemical analysis, and the relationship between TRPC1 and clinicopathological features of the patients was analyzed. The gastric carcinoma cells were transfected with siRNA-TRPC1, and the transfection efficiency of TRPC1 was detected by RT-PCR and Western blotting. Meanwhile, cell proliferation was detected by methyl thiazolyl tetrazolium (MTT). Moreover, superoxide dismutase (SOD), malondialdehyde and reactive oxygen species activity were detected by biochemical kit. The apoptotic level was detected by flow cytometry.Re? sults Compared with adjacent tissues, the expression of TRPC1 was significantly increased in tissues of gastric carcinoma (79.6% vs. 20.4%, P <0.05). The expression of TRPC1 in gastric cancer tissues was correlated with tumor TNM staging, differentiation degree andlymph node metastasis (P<0.05). Compared with the control group, TRPC1 mRNA and protein levels in the transfection group were significantly lower (P<0.05). In addition, the proliferation activity of siRNA-TRPC1 transfected cells was significantly decreased (P<0.05).Compared with the control group, malondialdehyde and reactive oxygen species levels increased while SOD activity decreased in siRNA-TRPC1 transfected cells (P <0.05); and the apoptotic level of siRNA-TRPC1 transfected cells increased significantly (P<0.05). Conclusions The expression of TRPC1 is up-regulated in gastric carcinoma tissues. Moreover, inhibition of TRPC1 expression bysiRNA increases oxidative stress and inhibits the cell proliferation.