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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 effect of miR-582-3p on the proliferation, migration and apoptosis of gastric cancer cell line SGC-7901. Methods: The expression of miR-582-3p in gastric epithelial cells (EGS-1) and gastric cancer cells (SGC-7901, MKN28) was detected by RT-PCR and miR-582-3p cells were selected. The SGC-7901 cells were transfected with miR-582-3p inhibitor. The cells were divided into blank control group, RPMI 1640 + SGC-7901 cells and RPMI 1640 + miR-582-3p + SGC-7901 cells. RPMI 1640 + miR-582-3p inhibitor + SGC-7901 cells. The proliferation and migration of cells were detected by MTT assay. The expression of miR-582-3p mRNA was detected by RT-PCR and the effect of miR-582-3p on the cell cycle and apoptosis of SGC-7901 cells by flow cytometry . Results: The highest expression of SGC-7901 was found in the three cell lines of EGS-1, MKN28 and SGC-7901 by miR-582-3p. SGC-7901 gastric cancer cells were selected for follow-up experiments. MTT showed that the expression of SGC- (P <0.05). The level of miR-582-3p mRNA in the experimental group was significantly lower than that in the control group (P <0.05). The cell cycle and apoptosis assay showed that the cell cycle of SGC-7901 cells in the experimental group was obvious (P <0.05). After 48 hours, the cells in the experimental group showed obvious apoptosis (P <0.05). Transwell experiments showed that the down-regulation of the expression of miR-582-3p could inhibit the invasion of SGC-7901 cells migrate. Conclusion: Inhibition of miR-582-3p can inhibit the proliferation, invasion and migration of gastric cancer cell SGC-7901, suggesting that miR-582-3p may play an important role in the development of gastric cancer.