NDRG2基因过表达影响卵巢癌细胞生物学功能的研究

Study on the effect of NDRG2 gene overexpression on the biological function of ovarian cancer cells

目的构建N-myc下游调节基因2(NDRG2)过表达质粒,研究NDRG2基因影响卵巢癌细胞增殖、凋亡及侵袭等生物学行为的作用.方法NDRG2过表达质粒载体感染卵巢癌细胞,从而实现NDRG2基因的在卵巢癌细胞中的过表达.之后通过CCK8试验观察卵巢癌细胞的生存能力及平板克隆形成试验观察卵巢癌细胞的增殖能力.流式细胞术检测细胞凋亡和细胞周期.结果质粒转染后,Q-PCR检测NDRG2基因特异过表达.Q-PCR试验观察NDRG2过表达质粒瞬时转染后NDRG2基因表达情况,与空载体组相比,NDRG2过表达转染组卵巢癌细胞NDRG2基因表达水平明显升高(OVCAR-3、SKOV3、CAOV3卵巢癌细胞t值分别为48.000、28.667、29.784,均P<0.05).CCK8检测、平板克隆试验结果显示过表达NDRG2基因显著抑制OVCAR-3、SKOV3及CAOV3细胞生长(t=22.928、13.797、10.332;χ^(2)=5.530、4.060、4.230,P<0.05),并促使上述3种卵巢癌细胞周期停滞于G1期.结论通过真核表达载体使细胞NDRG2基因的过表达,可以明显抑制OVCAR-3、SKOV3及CAOV3细胞的生长和增殖.

Objective To construct eukaryotic expression vector of n-myc downstream regulatory gene 2(NDRG2)overexpression,and to study the relationship between NDRG2 gene and biological behaviors such as proliferation,apoptosis and invasion of ovarian cancer.Methods NDRG2 overexpression vector was used to infect ovarian cancer cells,so as to realize the overexpression of NDRG2 gene in ovarian cancer cells.The effects of NDRG2 gene overexpression on the biological characteristics of ovcar-3,SKOV3,CAOV3 and other ovarian cancer cells were observed by plate cloning formation experiment and CCK8 experiment.Cell apoptosis and cell cycle were detected by flow cytometry.Results After plasmid transfection,Q-PCR was used to detect the specific overexpression of NDRG2 gene.Q-PCR results showed that compared with empty vector group,the expression levels of NDRG2 gene in ovarian cancer OVCAR-3 cells,ovarian cancer SKOV3 cells,and ovarian cancer CAOV3 cells in NDRG2 overexpression transfection group increased significantly(t=48.000,28.667,29.784,P<0.05).The results of CCK8 detection and plate cloning experiments showed that the overexpression of NDRG2 gene significantly inhibited the cell growth and tumor formation of ovcar-3,SKOV3 and CAOV3(t=22.928,13.797,10.332;χ^(2)=5.530,4.060,4.230,P<0.05),and promoted the cell cycle stagnation of the above three ovarian cancers in G1 phase.Conclusion The overexpression of NDRG2 gene by eukaryotic expression vector can significantly inhibit the growth and proliferation of ovcar-3,SKOV3 and CAOV3 cells.