Mda-7/IL-24基因转染对肝癌细胞凋亡影响的研究

目的 为了研究Mda-7／IL-24基因对肝癌细胞系Hep3B凋亡的影响。方法 构建了Mda-7／IL-24基因的真核表达载体pcDNA3．1／Mda-7／IL-24，用脂质体介导的基因转染法分别将真核重组体pcDNA3．1／Mda-7／IL-24和pcDNA3．1空载体质粒导人人肝癌细胞系Hep3B细胞中，经G418筛选获得细胞克隆。通过聚合酶链式反应（PCR）、免疫组织化学等方法检测基因和蛋白的表达，同时检测了细胞的生长和凋亡情况。结果 pcDNA3．1／Mda-7／IL-24转染的Hep3B细胞能够检测到Mda-7／IL-24的表达，可以抑制细胞生长，DNA琼脂糖凝胶电泳显示凋亡细胞特有的“梯状”条带，转染空载体和未转染的Hep3B细胞未见凋亡表现。结论 将外源性Mda-7／IL-24基因导人Hep3B细胞后，可以促进凋亡，Mda-7／IL-24作为一种肝癌的治疗基因，具有广泛的应用前景。

Effects of Mda-7/IL-24 gene transfection on growth of hepatocellular carcinoma cells

Objective To study the effects of Mda-7/IL-24 gene transfection on growth of hepatocellular carcinoma cells. Methods The mammalian expression recombinant pcDNA3. 1/Mda-7/IL- 24 vector was constructed. Then it and pcDNA3. 1 plasmid were respectively transfected into the HCC Hep3B cells. G418 was used to screen the transformants. The expression of Mda-7/IL-24 gene and protein was determined by RT-PCR, immunohistochemistry and Western blotting. The cell growth and the apoptotic effect were also studied. Results The expression of Mda-7/IL-24 protein was detected in the Hep3B cells transfected by pcDNA3. 1/Mda-7/IL-24. The growth rate and the apoptotic index of G418 resistant Hep3B cells with the expression of Mda-7/IL-24 protein was faster than that of vector transfected Hep3B cells and untransfected Hep3B cells. Conclusions Mda-7/IL-24 gene can inhibit the growth of HCC cells and can he used as a good candidate of tumor suppressor gene.