miR-214对HepG2细胞增殖及细胞周期影响

发布时间：2018-10-10 19:06

【摘要】：目的探讨miR-214对Hep G2细胞增殖及细胞周期的影响。方法 Realtime-PCR法检测肝癌细胞株SMMC-7721、Hep G2、SK-Hep-1、Huh 7、Hep3B及正常肝细胞L-02中miR-214表达量,并利用脂质体转染miR-214NC及miR-214 mimics,采用Realtime-PCR法检测转染效果;采用MTT法、流式细胞术分别检测miR-214对肝癌细胞活力、凋亡及细胞周期影响;蛋白印迹(WB)检测miR-214对肝癌细胞中细胞周期蛋白D1(cyclin D1),细胞周期蛋白依赖性激酶4(CDK4),生存素(survivin)及增值细胞核抗原(PCNA)表达影响。结果 miR-214在肝癌细胞SMMC-7721、SK-Hep-1、Huh 7、Hep3B、Hep G2中表达量[分别为(1.25±0.10)、(1.43±0.10)、(0.95±0.09)、(0.98±0.01)、(0.82±0.08)]明显低于正常肝细胞L-02(2.52±0.23),其中Hep G2中miR-214表达量最低。miR-214 mimics组Hep G2细胞中miR-214表达量(2.38±0.23)明显高于miR-214 NC组(0.83±0.08),miR-214mimics组Hep G2细胞活力(0.37±0.03)明显低于miR-214 NC组(0.78±0.07);与miR-214 NC组比较,miR-214mimics组Hep G2细胞凋亡率明显升高,细胞周期阻滞于G1期,cyclin D1、CDK4、survivin及PCNA表达量下调,差异均有统计学意义(P0.01)。结论上调miR-214表达可抑制肝癌细胞增殖,其机制可能与调控细胞周期相关蛋白表达水平有关。
[Abstract]:Objective to investigate the effect of miR-214 on the proliferation and cell cycle of Hep G2 cells. Methods Realtime-PCR assay was used to detect the expression of miR-214 in hepatoma cell line SMMC-7721,Hep G2SK-Hep-1Huh7Hep3B and normal hepatocytes L-02, and the transfection effect was detected by miR-214NC and miR-214 mimics, transfection with liposome, and the viability of hepatocarcinoma cells was detected by MTT assay and flow cytometry, respectively. The effects of miR-214 on the expression of cyclin D1 (cyclin D1), cyclin dependent kinase 4 (CDK4), survivin (survivin) and proliferating cell nuclear antigen (PCNA) were detected by Western blot (WB). Results the expression of miR-214 in hepatoma cell line (1.25 卤0.10), (1.43 卤0.10), (0.95 卤0.09), (0.98 卤0.01), (0.82 卤0.08) was significantly lower than that in normal hepatocyte L-02 (2.52 卤0.23). The expression of miR-214 in miR-214 mimics group was significantly higher than that in miR-214 NC group (2.38 卤0.23), and the activity of Hep G2 in miR-214mimics group was significantly higher than that in miR-214 NC group. (0.37 卤0.03) was significantly lower than that in miR-214 NC group (0.78 卤0.07), and the apoptosis rate of Hep G2 cells in miR-214mimics group was significantly higher than that in miR-214 NC group. The cell cycle was blocked in G1 phase, and the expression of cyclin D1 CDK4 and PCNA was down-regulated (P0.01). Conclusion Up-regulation of miR-214 expression can inhibit the proliferation of hepatoma cells, and its mechanism may be related to the regulation of cell cycle related protein expression level.
【作者单位】：牡丹江医学院第二附属医院肝胆外科;
【分类号】：R735.7

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