miR-582-3p对胃癌细胞SGC-7901增殖、迁移和凋亡的影响

Effect of miR-582-3p on proliferation,migration and apoptosis of gastric cancer SGC-7901 cells in vitro

目的:探讨miR-582-3p对胃癌细胞SGC-7901增殖、迁移和凋亡的影响。方法:RT-PCR检测miR-582-3p在胃上皮细胞(EGS-1)、胃癌细胞(SGC-7901、MKN28)中的表达,选出最高表达miR-582-3p细胞。合成miR-582-3p inhibitor转染SGC-7901细胞,实验分组,空白对照组:RPMI 1640+SGC-7901细胞,阴性对照组:RPMI 1640+miR-582-3p+SGC-7901细胞,实验组:RPMI 1640+miR-582-3p inhibitor+SGC-7901细胞。MTT检测细胞的增殖活性,Transwell检测增殖及迁移,RT-PCR法检测细胞miR-582-3p mRNA的表达水平,流式细胞仪检测miR-582-3p对SGC-7901细胞周期和凋亡的影响。结果:miR-582-3p在EGS-1、MKN28、SGC-7901三种细胞中SGC-7901表达最高,选取胃癌细胞SGC-7901进行后续实验,MTT显示实验组第24 h及48 h明显受到抑制,差异有统计学意义(P<0.05),RT-PCR提示实验组miR-582-3p mRNA水平降低(P<0.05),细胞周期和凋亡实验证明实验组24小时SGC-7901细胞周期出现明显阻滞,停滞在G_2/M期(P<0.05),第48小时实验组细胞出现明显凋亡(P<0.05),Transwell实验表明下调miR-582-3p的表达能够抑制SGC-7901细胞侵袭与迁移。结论:抑制miR-582-3p能抑制胃癌细胞SGC-7901的增殖、侵袭迁移,提示miR-582-3p在胃癌的发生、发展中可能发挥重要作用。

Objective：To investigate the influence of down - regulating miR -582 -3p effect on the proliferation, migration and apoptosis on gastric cancer SGC -7901 cells. Methods ： Human gastric cancer SGC -7901 ,MKN28 cell and gastric epithelial cell EGS -1 were detectied by RT -PCR,miR -582 -3p to detecteded the highest expression of miR -582 -3p cells. Synthesis of miR -582 -3p inhibitor transfection SGC -7901 cells, the experiment was di-vided into 3 groups ： Blank control group RPMI 1640 ＋ SGC - 7901 cells. Negative control group RPMI 1640 ＋ miR - 582 - 3p ＋ SGC - 7901 cells. Experimental group RPMI 1640 ＋ miR - 582 - 3p inhibitor of SGC - 7901 cells. Deter-mined by MTT test cell proliferation activity, RT - PCR method to detect cell miR -582 -3p mRNA expression level. Flow cytometry detection of miR - 582 - 3p in SGC - 7901 cell cycle and apoptosis. Results ： In three kinds of EGS - 1,SGC -7901 and MKN28 cells,SGC -7901 expression miR -582 - 3p was highest,choose gastric cancer SGC - 7901 cells for subsequent experiments,divided three group：Blank control group RPMI 1640 ＋ SGC -7901 cells. Neg-ative control group RPMI 1640 ＋ miR -582 -3p ＋ SGC -7901 cells. Experimental group RPMI 1640 ＋ miR -582 - 3p inhibitor of SGC - 7901 cells. MTT assay showed that the experimental group 24 h and 48 h significantly sup-pressed, the difference was statistically significant （P 〈0.05） ,RT - PCR indicate experimental group miR -582 -3p mRNA levels were lower （P 〈0. 05） ,cell cycle and apoptosis assay indicate experimental group 24 hours appearent blockage,increased in the G2/M phase proportion （P 〈0. 05） ,48 hours cells appeared apoptosis significantly （P 〈0. 05）. Transwell assay showed cell invation and migration was decreased in miR - 582 - 3p down regulated SGC - 7901 cells. Conclusion ：Inhibits miR -582 -3p expression can inhibit gastric cancer SGC -7901 proliferation, inva-sive and migration,miR -582 -3p may play an important role in gastric carcinogesis.