【摘要】 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。更多还原

【Abstract】 Objective To investigate the expression of miR-129-5 p 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-129-5 p 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-129-5 p in the gastric cacer tissues,the patients were divided into the high miR-129-5 p expression group and low miR-129-5 p expression group,and 1-year overall survival rates of the two groups were compared. The expression of miR-129-5 p 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-129-5 p mimic,miR-129-5 p inhibitor,and miR-129-5 p negative control sequence scrumble were transfected into the BGC-823 cell line,which were taken as the miR-129-5 p mimic group,miR-129-5 p 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-129-5 p in the gastric cancer tissues was lower than that in the adjacent tissues( P < 0. 01). The relative expression of miR-129-5 p 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-129-5 p 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-129-5 p expression group was lower than that of the high miR-129-5 p expression group( P < 0. 001). At 24 h after transfection,the OD values of the miR-129-5 p mimic group,miR-NC and miR-129-5 p 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-129-5 p mimic group,miR-NC group,and miR-129-5 p 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-129-5 p mimic group,miR-NC group,and miR-129-5 p inhibitor group,respectively,with statistically significant difference between three groups( all P < 0. 01). Conclusions The expression of miR-129-5 p is down-regulated in the gastric cancer tissues and cell lines. MiR-129-5 p could inhibit the proliferation and decrease the invasion of gastric cancer cells.更多还原