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Received:May 12, 2021 Published Online:September 20, 2021
Received:May 12, 2021 Published Online:September 20, 2021
中文摘要: 目的 探讨硫氧还蛋白还原酶1（TrxR1）在胃癌中的表达及对患者预后的影响，并研究沉默TrxR1对胃癌细胞增殖及凋亡的影响。方法 收集江苏大学附属句容医院2015年5月至2018年5月80例胃癌患者手术切除标本，通过免疫组化实验分析肿瘤组织及癌旁组织中TrxR1的表达。分析TrxR1的表达与病理分级、分期及组织学分化的关系，并结合随访数据分析与患者预后的关系。实时定量PCR和Western blot检测外购的正常胃黏膜细胞GES-1与人胃癌细胞AGS中TrxR1的表达。用小干扰RNA（siRNA）-TrxR1和siRNA-阴性对照(NC)转染AGS细胞，用实时定量PCR和Western blot、CCK-8实验、流式细胞术、划痕实验、Transwell实验对比siRNA转染对两种AGS细胞TrxR1 mRNA和蛋白表达水平及增殖、凋亡、迁移和侵袭的影响。结果 80例胃癌患者组织标本中，有52例（65%）TrxR1在胃癌组织中高表达，28例（35%）TrxR1在癌旁组织中高表达，结合随访资料，TrxR1的表达水平与肿瘤浸润深度、TNM分期及组织学分化程度及不良预后有关（P<0.05，P<0.01），而与性别、年龄、淋巴结转移、远处转移无关（P>0.05）。实时定量PCR和Western blot证实TrxR1在AGS细胞中相对于GES-1细胞中高表达（P<0.01）。转染后，siRNA-TrxR1组AGS细胞的TrxR1蛋白和mRNA表达水平较siRNA-NC组显著降低（P<0.01）;细胞的增殖能力较siRNA-NC组显著降低（P<0.01）。siRNA-TrxR1组与siRNA-NC组AGS细胞的迁移、侵袭能力差异无统计学意义（P>0.05）。siRNA-TrxR1组AGS细胞的凋亡率为（53.95±0.31）%，高于siRNA-NC组的（5.79±0.14）%（P<0.01）。结论 TrxR1在胃癌组织中高表达，且与患者的不良预后相关，siRNA可下调TrxR1的表达进而抑制胃癌细胞的增殖，促进细胞的凋亡。TrxR1可能成为胃癌靶向治疗的潜在靶点。
Abstract:Objective To investigate the expression of thioredoxin reductase 1 (TrxR1) in gastric cancer tissues and its prognostic effect and the influences of silencing TrxR1 on the proliferation and apoptosis of gastric cancer cells. Methods Gastric cancer and adjacent tissues paired samples of 80 patients were collected from May 2015 to May 2018 to analyze the expression of TrxR1 in tumor tissues and adjacent tissues by immunohistochemical assay. The associations of TrxR1 expression with pathological grade, stage and histological differentiation were analyzed, and the relationship between TrxR1 expression and prognosis was also analyzed in combination with follow-up data. The expressions of TrxR1 in normal gastric mucosa epithelial cells line GES-1 and gastric cancer cells AGS were detected by real-time quantitative PCR (RT-qPCR) and western blot. Small interfering RNA (siRNA) negative control (siRNA-NC) group and siRNA-TrxR1 group was constructed in AGS cells by siRNA transfection. RT-qRCR, western blot, CCK-8 assay, flow cytometry, scratch assay and transwell assay were used to analyze the effect of siRNA transfection on TrxR1 mRNA and protein expression level in siRNA-NC group and siRNA-TrxR1 group, and its effect on the proliferation, apoptosis, migration and invasion of AGS cells. Results TrxR1 were highly expressed in 52 cases (65%) of gastric cancer tissue samples and in 28 cases (35%) of adjacent tissues. In combination with the follow-up data, the expression level of TrxR1 was correlated with the depth of tumor invasion, stage, histological differentiation and poor prognosis (P<0.05) and was not correlated with gender, age, lymph node metastasis and distant metastasis (P>0.05). RT-qPCR and western blot confirmed that TrxR1 expression in AGS cells was significantly higher than that in GES-1 cells (P<0.01). After transfection, the expression levels of TrxR1 mRNA and protein in AGS cells in siRNA-TrxR1 group were significantly lower than those in siRNA-NC group (P<0.01) .The proliferation of AGS cells in siRNA-TrxR1 group was significantly lower than that in siRNA-NC group (P<0.01), and there was no significant difference in cell migration and invasion of AGS cells between two groups (P>0.05). Apoptosis rate of AGS cells in siRNA-TrxR1 group was higher than that in siRNA-NC group ［ (53.95±0.31) % vs（5.79±0.14) %,P<0.01］. Conclusion TrxR1 is highly expressed in gastric cancer tissues and is related to the poor prognosis of patients. Downregulation of TrxR1 could inhibit the proliferation of gastric cancer cells and promote cell apoptosis. TrxR1 may be a potential target for targeted therapy of gastric cancer.
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基金项目:江苏省干部保障科研项目（BJ16008）
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