miR-29a对人胃癌细胞增殖、迁移的影响及其机制研究

doi:10.3969/j.issn.1004-616x.2020.06.008

摘要/Abstract

摘要： 目的：探讨微小RNA-29a（miR-29a）对人胃癌细胞增殖、迁移的影响及其机制。方法：胃癌SGC-7901细胞随机分为miR-29a转染组、质粒对照组、空白对照组、信号转导子与转录激活子3（STAT3）抑制组。miR-29a转染组、质粒对照组采用脂质体转染法分别将has-miR-29a mimics质粒、siRNA对照质粒转染至人胃癌SGC-7901细胞，STAT3抑制组细胞加入STAT3抑制剂溶液，空白对照组不做处理。实时荧光定量聚合酶链反应（qPCR）和Western blot法检测转染后细胞中miR-29a、血管内皮生长因子（VEGF）、STAT3、细胞周期蛋白D1（cyclin D1）mRNA和蛋白的表达情况；MTT、划痕实验分别检测细胞增殖和迁移情况。结果：qPCR和Western blot实验结果显示，与空白对照组和质粒对照组比较，miR-29a转染组的miR-29a相对表达水平升高，VEGF、cyclin D1 mRNA及蛋白相对表达水平与p-STAT3/STAT3比值均下降（P < 0.01）。MTT实验与划痕实验结果显示，与空白对照组和质粒对照组比较，miR-29a转染组细胞在培养24、48、72 h的细胞相对增殖率及迁移率降低（P < 0.01）。与空白对照组比较，STAT3抑制组细胞的cyclin D1 mRNA及蛋白相对表达水平、p-STAT3/STAT3比值均降低，细胞相对增殖率与迁移率降低（P < 0.01）。结论：miR-29a能降低VEGF mRNA、蛋白的表达，抑制人胃癌细胞增殖和迁移，其机制可能与下调p-STAT3及cyclin D1的表达有关。

关键词: 微小RNA-29a, 人胃癌SGC-7901细胞, 增殖, 凋亡, 信号传导, 信号转导子与转录激活子3

Abstract: OBJECTIVE: To investigate the effect of microRNA-29a (miR-29a) on proliferation and migration of human gastric cancer cells in vitro. METHODS: Gastric cancer SGC-7901 cells were randomly divided into miR-29a transfection group,plasmid control group,blank control group,and signal transducers and activators of transcription 3 (STAT3) inhibition group. The miR-29a transfection and the plasmid control groups were transfected with the hsa miR-29a mimics plasmids and the siRNA control plasmids,respectively,by liposome transfection. The STAT3 inhibition group received the STAT3 inhibitor solution and the blank control group was not treated. Real-time fluorescent quantitative polymerase chain reaction (qPCR) and Western blot were used to detect expression of miR-29a,vascular endothelial growth factor (VEGF),STAT3,cyclin D1 mRNA and protein in the transfected cells. MTT and scratch tests were used to detect cell proliferation and migration. RESULTS: qPCR and Western blot results show that,compared with the blank control and the plasmid control groups,expression levels of miR-29a in the miR-29a transfection group was increased,and expression levels of VEGF,cyclin D1 mRNA and protein,p-STAT3/STAT3 were decreased (P < 0.01). MTT and scratch tests show that,compared with the blank control and the plasmid control groups,the miR-29a transfection group had lower cell proliferation and migration rates at 24,48 and 72 h (P < 0.01). After application of the STAT3 inhibitor,compared with the blank control group,expression levels of cyclin D1 mRNA and protein,p-STAT3/STAT3,and MTT test cell relative proliferation rates at 24,48,72 h were lower in the STAT3 inhibition group (P < 0.01). CONCLUSION: Our data show that MiR-29a reduced the expression of VEGF mRNA and protein and inhibited the proliferation and migration of human gastric cancer cells. A mechanism for these effects may be related to down-regulation of p-STAT3,and cyclin D1 mRNA and protein expression.

Key words: microRNA-29a, human gastric cancer SGC-7901 cell, proliferation, apoptosis, signal transduction, signal transducers and activators of transcription 3

中图分类号:

R735.2

刘鹏, 白意晓. miR-29a对人胃癌细胞增殖、迁移的影响及其机制研究[J]. 癌变·畸变·突变, 2020, 32(6): 452-456,463.

LIU Peng, BAI Yixiao. Effect of miR-29a on proliferation and migration of human gastric cancer cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2020, 32(6): 452-456,463.

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