miR-129-5p在胃癌组织、细胞系中表达及其对细胞增殖和侵袭能力的影响

Expression of miR-129-5p in gastric cancer tissues and cell lines and its effects on proliferation and invasion

目的检测微小RNA-129-5p(miR-129-5p)在胃癌组织及细胞系中的表达,并探讨其对细胞增殖和侵袭能力的影响。方法采用实时荧光定量PCR法检测70例胃癌术后患者癌及癌旁正常组织中miR-129-5p表达,分析其与患者临床病理特征和预后间的关系。根据胃癌组织中miR-129-5p的中位表达量,将患者分为miR-129-5p高表达组和miR-129-5p低表达组,比例两组的1年总生存率。采用实时荧光定量PCR法检测胃癌细胞系(SGC-7901、MKN-45和BGC-823)和正常胃黏膜细胞系(GES-1细胞)中miR-129-5p表达。使用Lipofectamine3000将miR-129-5p模拟物、miR-129-5p抑制剂及miR-129-5p阴性对照序列Scramble转染对数期BGC-823细胞系,并分成miR-129-5p mimic组、miR-129-5p inhibitor组及miR-NC组。采用CCK-8法及Transwell实验分别测定细胞增殖及侵袭能力。结果胃癌组织中miR-129-5p相对表达量低于癌旁正常组织(P<0.01);胃癌细胞系(MKN-45、BGC-823和SGC-7901)中miR-129-5p相对表达量低于GES-1细胞系(P均<0.01)。miR-129-5p表达与肿瘤大小和淋巴结转移有关(P均<0.05),与患者年龄﹑性别﹑组织分化程度及TNM分期无关(P均﹥0.05)。miR-129-5p低表达组1年总生存率低于miR-129-5p高表达组(P<0.001)。转染后24 h,miR-129-5p mimic组、miR-NC组、miR-129-5p inhibitor组OD值分别为1.03±0.03、1.93±0.05、2.25±0.08,三组比较,P<0.01;转染后48 h的OD值分别为1.22±0.03、2.31±0.06、3.02±0.18,三组比较,P<0.01;转染后72 h的OD值分别为2.25±0.03、3.11±0.06、4.02±0.15,三组比较,P<0.01。细胞集落形成实验结果显示,miR-129-5p mimic组、miR-NC组、miR-129-5p inhibitor组细胞集落数目分别为(42±5)、(68±7)、(210±13)个,三组比较,P<0.01。Transwell实验结果显示,miR-129-5p mimic组、miR-NC组、miR-129-5p inhibitor组侵袭细胞数分别为(20±3)、(48±5)、(82±3)个,三组比较,P<0.01。结论miR-129-5p在胃癌组织及细胞系中表达均下调,miR-129-5p可抑制胃癌细胞增殖,降低细胞侵袭能力。

Objective To investigate the expression of miR-129-5p in the gastric cancer tissues and cell lines of gastric cancer patients,and to explore its effects on cell proliferation and invasion.Methods qRT-PCR was used to detect the expression of miR-129-5p in the gastric cancer and adjacent tissues of 70 patients with gastric cancer.The relationship between the expression and clinicopathological characteristics and prognosis was analyzed.According to the median expression of miR-129-5p in the gastric cacer tissues,the patients were divided into the high miR-129-5p expression group and low miR-129-5p expression group,and 1-year overall survival rates of the two groups were compared.The expression of miR-129-5p in the gastric cancer cell lines(SGC-7901,MKN-45,and BGC-823)and normal gastric mucosa cell line(GES-1)was compared.By using Lipofectamine 3000,miR-129-5p mimic,miR-129-5p inhibitor,and miR-129-5p negative control sequence scrumble were transfected into the BGC-823 cell line,which were taken as the miR-129-5p mimic group,miR-129-5p inhibitor group,and miR-NC group,respectively.CCK-8 and Transwell assay were used to detect the cell proliferation and invasion abilities.Results The relative expression of miR-129-5p in the gastric cancer tissues was lower than that in the adjacent tissues(P<0.01).The relative expression of miR-129-5p in the gastric cancer cell lines(MKN-45,BGC-823 and SGC-7901)was lower than that in cell line GES-1(P<0.01).The expression of miR-129-5p was related to tumor size and lymph node metastasis(both P<0.05),but not to age,sex,differentiation degree or TNM stage(all P>0.05).The 1-year overall survival rate of the low miR-129-5p expression group was lower than that of the high miR-129-5p expression group(P<0.001).At 24 h after transfection,the OD values of the miR-129-5p mimic group,miR-NC and miR-129-5p inhibitor group were 1.03±0.03,1.93±0.05 and 2.25±0.08,respectively,with statistically significant difference between three groups(all P<0.01);at 48 h after transfection,the OD values of three groups were 1.22±0.03,2.31±0.06,and 3.02±0.18,respectively,with statistically significant difference between three groups(all P<0.01).At 48 h after transfection,the OD values of three groups were 2.25±0.03,3.11±0.06,and 4.02±0.15,respectively,with statistically significant difference between three groups(all P<0.01).The number of cell colonies in the miR-129-5p mimic group,miR-NC group,and miR-129-5p inhibitor group were 42±5,68±7 and 210±13,respectively,with statistically significant difference between three groups(all P<0.01).Transwell assay showed that there were 20±3,48±5,and 82±3 invasive cells in the miR-129-5p mimic group,miR-NC group,and miR-129-5p inhibitor group,respectively,with statistically significant difference between three groups(all P<0.01).Conclusions The expression of miR-129-5p is down-regulated in the gastric cancer tissues and cell lines.MiR-129-5p could inhibit the proliferation and decrease the invasion of gastric cancer cells.