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谭立业, 李立德, 汪晶, 刘卓. miR-214对HepG2细胞增殖及细胞周期影响[J]. 中国公共卫生, 2017, 33(4): 614-617. DOI: 10.11847/zgggws2017-33-04-25
引用本文: 谭立业, 李立德, 汪晶, 刘卓. miR-214对HepG2细胞增殖及细胞周期影响[J]. 中国公共卫生, 2017, 33(4): 614-617. DOI: 10.11847/zgggws2017-33-04-25
TAN Li-ye, LI Li-de, WANG Jing.et al, . Effect of miR-214 on proliferation and cell cycle in hepatocellular carcinoma cells[J]. Chinese Journal of Public Health, 2017, 33(4): 614-617. DOI: 10.11847/zgggws2017-33-04-25
Citation: TAN Li-ye, LI Li-de, WANG Jing.et al, . Effect of miR-214 on proliferation and cell cycle in hepatocellular carcinoma cells[J]. Chinese Journal of Public Health, 2017, 33(4): 614-617. DOI: 10.11847/zgggws2017-33-04-25

miR-214对HepG2细胞增殖及细胞周期影响

Effect of miR-214 on proliferation and cell cycle in hepatocellular carcinoma cells

  • 摘要: 目的 探讨miR-214对HepG2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、HepG2、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-214 NC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(cyclinD1),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、HepG2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HepG2中miR-214表达量最低。miR-214 mimics组HepG2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-214 mimics组HepG2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-214 mimics组HepG2细胞凋亡率明显升高,细胞周期阻滞于G1期,cyclinD1、CDK4、survivin及PCNA表达量下调,差异均有统计学意义(P<0.01)。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 explore inhibitive effect of miR-214 on proliferation and cell cycle in hepatocellular carcinoma cells.Methods The expression of miR-214 in hepatocellular carcinoma cells of SMMC-7721,HepG2,SK-Hep-1,Huh 7,and Hep3B and normal liver cell L-02 was detected with realtime-PCR.Hepatocellular carcinoma cells were transfected with miR-214 NC and miR-214 mimics by liposome and the outcome of the transfection was determined with realtime-PCR.The cell viability,cell apoptosis and cell cycle was assessed with (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay and flow cytometry,respectively.The expression of cyclin D1,cyclin-dependent protein kinases (CDK4),survivin,and proliferating cell nuclear antigen (PCNA) were detected with Western blot.Results The expression of miR-214 in the cells of SMMC-7721 (1.25±0.10),SK-Hep-1 (1.43±0.10),Huh 7 (0.95±0.09),Hep3B (0.98±0.0),and HepG2 (0.82±0.08) were all lower than that in L-02 cells (2.52±0.23) and the expression of miR-214 was the lowest in HepG2 cells.The expression of miR-214 in miR-214 mimics HepG2 cells (2.38±0.23) was higher than that in miR-214 NC HepG2 cells (0.83±0.08).The cell viability of miR-214 mimics HepG2 cells (0.37±0.03) was lower than that of miR-214 NC cells (0.78±0.07).Compared with those of the miR-214 NC HepG2 cells,the cell apoptotic rate was increased;cell cycle was arrested at G1;and expressions of cyclin D1,CDK4,survivin,and PCNA were down-regulated in miR-214 mimics HepG2 cells,with significant differences between the two groups (P<0.01 for all).Conclusion miR-214 mimics inhibits HepG2 cell proliferation;the mechanism of the inhibitive effect may be related to the expression of cell cycle regulation related protein.

     

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