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目的研究X-染色体相关凋亡抑制蛋白(XIAP)相关因子1(XAF1)对肝癌细胞增殖和凋亡的影响。方法构建正义pcDNA3～XAF1和反义pcDNA3-XAF1-AS质粒,通过脂质体转染技术将质粒DNA转入SMMC7721肝癌细胞株并建立稳定高表达XAF1的肝癌细胞克隆。应用MTT、流式细胞仪、TUNEL法检测肝癌细胞的增殖和凋亡；应用RT-PCR和Western印迹法检测基因在mRNA和蛋白水平的表达。结果XAF1在SMMC7721肝癌细胞株中有表达,但低于正常肝脏细胞水平。稳定高表达XAF1基因可抑制肝癌细胞的增殖和诱导凋亡,并能增加其对化疗药物5-氟尿嘧啶和羟基喜树碱的敏感性。结论XAF1能抑制肝癌细胞增殖和诱导其凋亡。
Objective To investigate the effect of XAF1 on the proliferation and apoptosis of hepatocellular carcinoma cells. Methods Plasmids pcDNA3 ~ XAF1 and antisense pcDNA3-XAF1-AS were constructed and transfected into SMMC7721 hepatoma cell line by lipofection technique. Clone of hepatoma cells stably expressing XAF1 was established. The proliferation and apoptosis of hepatocellular carcinoma cells were detected by MTT, flow cytometry and TUNEL. The mRNA and protein levels were detected by RT-PCR and Western blot. Results XAF1 was expressed in SMMC7721 hepatoma cell lines, but lower than normal liver cells. Stably overexpressing XAF1 gene can inhibit the proliferation and induce apoptosis of hepatoma cells and increase its sensitivity to chemotherapeutic agents 5-fluorouracil and hydroxycamptothecin. Conclusion XAF1 can inhibit the proliferation and induce the apoptosis of hepatoma cells.