nm23-H1对顺铂化疗效果影响的体外研究

The study of increased sensitivity to cisplatin by nm23-H1 in vitro

目的nm23-H1对顺铂化疗效果的影响及其可能机制。方法通过脂质体转染法,将nm23-H1转染入舌癌细胞株Tca8113,经免疫组化和Western-bloting鉴定,建立稳定表达细胞株。检测转染组和未转染组顺铂杀伤率、细胞凋亡和线粒体膜电位的变化。结果nm23-H1过表达可以明显提高顺铂对舌癌细胞的杀伤率,使细胞凋亡增强,线粒体膜电位降低。这种作用可以被哇巴因抑制。结论nm23-H1可提高顺铂对舌癌细胞化疗的敏感性,其可能机制是nm23-H1降低线粒体膜电位,顺铂进入细胞内增加,导致细胞凋亡和坏死。

Objective To study the mechanism of increased sensitivity to cisplatin by nm23-H1. Methods The plasmid, pCMV-BamH1-Neo-nm23-H1, was transfected to cell line Tca8113 by lipofection strategy and was selected by G418 for 5 weeks. The stable products of nm23-H1 gene were identified by Immunohistofluorescence and Western-blotting. Using MTT and flow cytometer, we detected the changes of cell mortality rate, apoptosis and mitochondrial membrane potential. Results We successfully established the cell line of stable overexpression of nm23-H1. In vitro, the cell mortality rate and apoptosis were increased significantly in stable expression cell line of nm23-H1, compared with those in Tca8113 cell line of non-transfection; this effect could be inhibited by oubain which is an inhibitor of Na（＋）/K（＋）-ATPase. Mitochondrial membrane potential was decreased significantly in stable expression cell line of nm23-H1. Conclusion Nm23-H1 can increase the sensitivity of cisplatin on Tca8113 cell line; The mechanism may be the mitochondrial membrane potential is decreased by nm23-H1 so that intracellular platinum are increased to finally cause the apoptosis and necrosis of cells.