下调肝癌衍生生长因子表达水平对胶质瘤U373细胞增殖及凋亡的影响

Effects of down regulating hepatoma derived growth factor of glioma on the proliferation and apoptosis of U373 cells

摘要:

目的：研究下调肝癌衍生生长因子(Hepatoma-derived growth factor,HDGF)的表达水平对胶质瘤细胞U373增殖及凋亡能力的影响。方法：设计并合成HDGF靶向siRNA，以脂质体为介质瞬时转染入U373细胞，采用RT-PCR及Western blot检测HDGF基因及蛋白表达水平的变化，判断RNA干扰的效果、流式及MTT法检测细胞凋亡及增殖变化。结果：用脂质体瞬时转染siRNA至U373细胞中，RT-PCR及Western blot实验结果证实转染48 h后，U373细胞中HDGF基因及蛋白表达水平明显下调。MTT 结果显示，RNAi下调HDGF 后的U373细胞相对于空白和阴性对照组，细胞在体外增殖能力明显减弱（P<0.01）,U373细胞凋亡率明显上升（P<0.05）。结论：HDGF蛋白在胶质瘤的发生发展过程中起着促进作用，可能成为一个新的肿瘤分子治疗靶点。

Abstract:

Objective:To evaluate the effects of down regulating hepatoma-derived growth factor (HDGF) of glioma on the proliferation and apoptosis of U373 cells.Methods:siRNA targeting human HDGF gene was designed and transfected transiently into U373 cells by lipofectamine. RT-PCR and Western blot were used to detect the changes in expressions of HDGF gene and protein as well as to evaluate the efficacy of RNA interference. Flow cytometric detection and MTT method were used to detect cell apoptosis and proliferation. Results:RT-PCR and Western blot results showed that the expressions of HDGF gene and protein declined obviously in U373 cells at 48 hours after the transfection. Meanwhile,the MTT results indicated that the proliferation in vitro was remarkably decreased and the apoptosis rate was significantly increased in U373 cells compared with those in blank and negative control group after down regulating HDGF expressions (P<0.01 andP<0.05). Conclusion:HDGF plays a conductive role in the development of glioma,which may become a new molecular marker in tumor diagnosis and treatment.