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目的:在肝癌细胞Hep G2中过表达外源NAIF1(核凋亡诱导因子1),探讨NAIF1的亚细胞定位以及对Hep G2增殖和迁移能力的影响。方法:以真核表达质粒p EGFP-N1为对照组,p EGFP-N1-NAIF1为实验组,瞬时转染肝癌细胞Hep G2,利用免疫印迹方法检测NAIF1蛋白表达效率;以DAPI染核,荧光显微镜下观察绿色荧光蛋白定位,确定NAIF1的亚细胞定位;通过MTT方法绘制细胞增殖曲线;通过transwell小室法检测NAIF1对Hep G2迁移能力的影响。结果:在肝癌细胞Hep G2中,外源表达NAIF1主要定位于细胞核;与对照组Hep G2/p EGFP-N1相比,Hep G2/p EGFP-N1-NAIF1的细胞增殖、迁移能力下降(P<0.05)。结论:外源表达NAIF1蛋白定位于Hep G2细胞核,过表达NAIF1抑制Hep G2的细胞增殖与迁移能力,NAIF1可能作为肝癌治疗的潜在靶点。
OBJECTIVE: To overexpress exogenous NAIF1 (nuclear factor of inducible factor 1) in hepatocellular carcinoma Hep G2 cells to investigate the subcellular location of NAIF1 and its effect on the proliferation and migration of Hep G2 cells. Methods: The eukaryotic expression plasmid pEGFP-N1 was used as a control group. P EGFP-N1-NAIF1 was used as an experimental group, transient transfected into Hep G2 cells, and the expression of NAIF1 protein was detected by immunoblotting. The localization of green fluorescent protein (EGFP) was observed to determine the subcellular localization of NAIF1. The cell proliferation curve was drawn by MTT assay. The effect of NAIF1 on the migration of Hep G2 was detected by transwell chamber assay. Results: The expression of NAIF1 mainly localized in the nucleus of Hep G2 cells. Compared with Hep G2 / EGFP-N1, the proliferation and migration of Hep G2 / EGFP-N1-NAIF1 cells decreased (P < 0.05). CONCLUSION: The exogenous NAIF1 protein is located in the nucleus of Hep G2 cells. Overexpression of NAIF1 can inhibit the proliferation and migration of Hep G2 cells. NAIF1 may be a potential therapeutic target for hepatocellular carcinoma.