沉默Nek2基因对卵巢癌SKOV3细胞周期的影响

Effect of silencing Nek2 gene on cell cycle of ovarian cancer SKOV3 cells

目的研究RNA干扰NIMA相关激酶2(Nek2)表达对卵巢癌细胞SKOV3细胞周期的影响及其相关的分子机制。方法将针对Nek2基因的siRNA转染至SKOV3细胞中,采用流式细胞技术检测SKOV3细胞周期变化,并用Western blot技术检测Nek2-siRNA转染入卵巢癌SKOV3细胞48h后,与细胞周期相关的因子cyclinB1、CDK1及P27蛋白表达水平以及ERK1/2磷酸化水平的变化。结果空白对照组、阴性对照组和RNA干扰组中处于G2/M期的细胞比例分别为13.72%、12.27%和1.56%,与两对照组比较,处于G2/M期的干扰组细胞明显减少(P<0.05)。Nek2基因沉默后,与两对照组比较,SKOV3细胞内cyclinB1和CDK1的蛋白表达水平明显下降,P27的蛋白表达水平明显上调,SKOV3细胞内ERK1/2磷酸化水平明显下降(P<0.05)。结论沉默Nek2基因,可阻止卵巢癌SKOV3细胞启动有丝分裂,从而抑制其增殖。

Objective To study the effect of silencing Nek2 via RNAi on cell cycle of ovarian cancer SKOV 3 cells and the re‐lated molecular mechanism .Methods The Nek2‐siRNA was transfected into the ovarian cancer SKOV3 cells .The change of cell cy‐cle of SKOV3 cells at 48 h after transfection was examined by the flow cytometry technique ;Western blot assay was used to deter‐mine the change of level of the cell cycle related factors cyclinB1 ,CDK1 ,P27 and the phosphorylation level of the ERK1/2 after Nek2‐siRNA transfection for 48 h .Results The flow cytometry detection results showed that the proportion of the cells in G 2/M stage in the blank control group ,negative control group and RNAi group was 13 .72% ,12 .27% and 1 .56% respectively .Compared with the control group ,the number of the cells in G2/M stage in the transfected group was reduced obviously （P〈 0 .05） .The Western blot detection results showed that compared with the control group ,the expression of cyclinB1 and CDK1 protein in SK‐OV3 cells was significantly reduced ,the expression of P27 was increased after silencing Nek2 and the phosphorylation level of ERK1/2 in SKOV3 cells was significantly reduced after silencing Nek2 gene（P〈 0 .05） .Conclusion Silencing Nek2 gene might block the ovarian cancer cell line SKOV 3 initiating mitosis ,thus inhibit their proliferation .