miR-181b-5p对胃癌细胞HGC-27增殖和凋亡的影响

The Effect of miRNA-181b-5p on Proliferation and Apoptosis of Human Gastric Cancer Cell HGC-27

该文检测了人胃癌血清和胃癌细胞HGC-27中miR-181b-5p的水平,并研究了miR-181b-5p对胃癌细胞HGC-27增殖和凋亡能力的影响,以探讨miR-181b-5p在胃癌发生和发展中可能的作用。荧光定量PCR(Real-time quantitative PCR,q RT-PCR)检测胃癌患者和健康人群血清中miR-181b-5p的水平以及胃癌细胞HGC-27和胃黏膜细胞GES-1中miR-181b-5p的水平,采用脂质体瞬时转染技术将miR-181b-5p抑制剂(inhibitors)以及inhibitors阴性对照分别转入胃癌细胞HGC-27中,用q RTPCR验证转染后miR-181b-5p的水平,CCK-8法检测miR-181b-5p对胃癌细胞HGC-27的增殖能力的影响,流式细胞术检测转染后胃癌细胞HGC-27细胞周期和凋亡的变化。q RT-PCR结果显示,胃癌患者血清中miR-181b-5p的水平高于健康人群(P<0.05);胃癌细胞HGC-27中miR-181b-5p的水平显著高于胃黏膜细胞GES-1中的水平(P<0.01);转染miR-181b-5p inhibitors后其miR-181b-5p的水平显著低于inhibitors阴性对照组(P<0.01)。CCK-8结果显示,将miR-181b-5p的水平下调后,胃癌细胞HGC-27的增殖能力明显下降(P<0.01)。流式细胞术检测结果显示,转染miR-181b-5p inhibitors后,胃癌细胞HGC-27的凋亡率显著增高(P<0.01)。细胞周期检测结果显示,转染miR-181b-5p inhibitors后,其S期明显缩短,G2/M期延长,细胞增殖受到抑制。以上结果表明,血清miR-181b-5p可能作为胃癌早期诊断的一个分子肿瘤标志物;miR-181b-5p在胃癌的发生和发展过程中可能起到癌基因的作用,并有可能作为胃癌分子治疗的一个新靶点。

The miR NA-181b-5p levels in serum of gastric cancer patients and gastric cancer cell HGC-27were detected,and the effect of miR NA-181b-5p on proliferation and apoptosis of gastric cancer cell HGC-27 were investigated,which in order to explore the role of miR NA-181b-5p in the progress and development of gastric cancer.Real-time quantitative PCR（qR T-PCR）was performed to detect miR NA-181b-5p levels in serum of gastric cancer patients and healthy people,and in gastric cancer cell HGC-27 and gastric mucosal cell GES-1.The miR NA-181b-5p inhibitiors and inhibitors negative control was tranfected into gastric cancer cell HGC-27 by Lipofectamine 2000,respectively.Then miR NA-181b-5p was identified by qR T-PCR.The effect of miR NA-181b-5p on the proliferation of gastric cancer cell HGC-27 was detected by CCK-8 assay.The cell apoptosis and cell cycle distribution were analyzed by flow cytometry.The q RT-PCR result showed that the miRNA-181b-5p level in serum of gastric cancer was increased compared with the healthy people（P〈0.05）,and was significantly increased in gastric cancer cell HGC-27compared with gastric mucosal cell GES-1（P〈0.01）.The miR NA-181b-5p level in gastric cancer cell was decreased after miR NA-181b-5p inhibitiors transfection compared with the inhibitors negative control（P〈0.01）.The result of CCK-8 assay showed that the proliferation of gastric cancer cell HGC-27 was significantly reduced while the miR NA-181b-5p level was reduced（P〈0.01）.The result of flow cytometry showed that the apoptosis of gastric cancer cell HGC-27 was significantly up-regulated after transfection（P〈0.01）,and the proportion of S-stage cells was decreased as well as the proportion of G2/M-stage cells was increased.The proliferation of gastric cancer cell was inhibited after the tranfection of miR NA-181b-5p inhibitors.These results indicated that the serum level of miR NA-181b-5p may act as an molecular marker for early diagnosis of gastric cancer,and miR NA-181b-5p may act as oncogene in the progress and development of gastric cancer and serve as a new target for the molecular treatment of gastric cancer.