硫氧还蛋白还原酶1(TrxR1)在胃癌中的表达及对胃癌细胞生长的影响

Expression of thioredoxin reductase 1(TrxR1)in gastric cancer and its effect on the growth of gastric cancer cells

目的:探讨胃癌组织硫氧还蛋白还原酶1(TrxR1)表达与生存时间的关系及其对胃癌细胞生长的影响。方法:用Real-time PCR法检测76例胃癌组织及癌旁TrxR1 mRNA表达,并分析其与胃癌患者临床病理特征及预后的关系;随机选取3例胃癌组织及癌旁组织,采用免疫组化法、Western blot法检测TrxR1蛋白表达。采用Western blot法和Real-time PCR法检测胃癌细胞系及人胃粘膜上皮细胞中TrxR1的表达。采用小RNA干扰序列(siRNA)处理AGS细胞,根据处理方法不同将AGS细胞分为3组:阴性对照组:转染NC-siRNA、TRXR1 siRNA干扰1组:转染TRXR1-siRNA1、TRXR1 siRNA干扰2组:转染TRXR1-siRNA2。使用Real-time PCR法检测各组AGS细胞中TrxR1 mRNA的表达,克隆形成试验和MTT法检测AGS细胞生长情况。结果:胃癌组织中TrxR1 mRNA和蛋白表达量均显著性上调,TrxR1主要定位于细胞质中。TrxR1高表达与患者TNM分期及淋巴结转移有关,且TrxR1高表达组患者的中位生存时间短于低表达组(P<0.05)。胃癌细胞中TrxR1表达量高于人胃粘膜上皮细胞系中的表达。TRXR1-siRNA1组AGS细胞和TRXR1-siRNA2组AGS细胞中TrxR1 mRNA和蛋白与NC-siRNA组相比均显著性降低(P<0.05),且AGS细胞克隆形成与增殖能力均降低(P<0.05)。结论:胃癌组织中TrxR1高表达提示患者预后不良,沉默TrxR1能抑制胃癌细胞的增殖。

Objective:To investigate the relationship between the expression of thioredoxin reductase 1(TrxR1)in gastric cancer tissues and the survival time of patients with gastric cancer and its effect on the growth of gastric cancer cells.Methods:The expression level of TrxR1 mRNA in gastric cancer tissues and adjacent tissues of 76 patients was determined by real-time PCR assays,and the relationship between the mRNA expression level of TrxR1 and the clinicopathological characteristics and prognosis of gastric cancer patients was analyzed.Three gastric cancer tissues and adjacent tissues were randomly selected,and the expression of TrxR1 protein was detected by immunohistochemistry and Western blot.The expression levels of TrxR1 in human gastric cancer cells line and gastric mucosa epithelial cells were determined by Western blot and quantitative real-time PCR assays.Then,AGS cells were transfected with siRNA sequences to silence the expression of TrxR1.AGS cells were divided into 3 groups according to different processing negative control group:transfected with NC-siRNA,TRXR1 siRNA interference group 1:transfected with TRXR1-siRNA1,TRXR1 siRNA interference 2 group:transfected with TRXR1-siRNA2.The expression of TrxR1 mRNA in AGS cells of each group was detected by Real-time PCR.The proliferation of AGS cells was determined by MTT and colony formation assays following TrxR1 silencing.Results:TrxR1 mRNA and protein expression levels in gastric cancer tissues were significantly up-regulated compared with adjacent tissues,and were mainly located in the cytoplasm.High expression of TrxR1 was associated with TNM stage and lymph node metastasis,and the overall survival time of patients with high expression of TrxR1 was shorter than those with low expression level.TrxR1 mRNA and protein in AGS cells of TRXR1-siRNA1 group and AGS cells of TRXR1-siRNA2 group were significantly reduced compared with NC-siRNA group(P<0.05).And AGS cell clone formation and proliferation ability were decreased.Conclusion:The high expression of TrxR1 in gastric cancer tissues indicates poor prognosis of patients,and the silencing of TrxR1 can inhibit the proliferation of gastric cancer cells.