重组腺病毒介导hTIMP-1对人肝癌细胞系体外增殖的影响

Effects of Recombinant Adenovirus-Mediated Overexpression of hTIMP-1 on Proliferation of Human Liver Cancer Cell Line HepG2 in Vitro

目的探讨人基质金属蛋白酶组织抑制因子-1(hTIMP-1)过表达对人肝癌细胞系HepG2体外增殖的作用。方法构建携载hTIMP-1全长cDNA的重组腺病毒载体,感染HepG2细胞,应用Western blot及RT-PCR方法检测TIMP-1蛋白及mRNA的表达,透射电镜观察细胞超微结构改变,MTT法检测hTIMP-1对HepG2细胞生长的影响。结果成功构建携载hTIMP-1的重组腺病毒载体,感染HepG2细胞,实现其体外稳定表达,hTIMP-1过表达对HepG2体外增殖有明显抑制作用。结论hTIMP-1过表达可抑制人肝癌细胞系HepG2的体外增殖,可用于肝癌基因治疗的研究。

Objective To explore the effects of overexpression of human tissue inhibitors of metalloproteinase-1 （hTIMP-1） on proliferation of human liver cancer cell line HepG2 in vitro. Methods A recombinant adenoviral vector containing full-length cDNA of hTIMP-1 was generated and transfected into HepG2. The viral titer was checked by measuring GFP, and the expression of hTIMP-1 in vitro was detected by the techniques of Western blot and semi-quantitative RT-PCR. The ultrastructure was observed by transmission electron microscope and the effects of overexpression of hTIMP-1 on proliferation of HepG2 in vitro was analyzed by MTT assay and growth curve. Results The resultant AdhTIMP-1 was successfully constructed and the expression of hTIMP-1 was detected by Western blot and RT-PCR. The growth and proliferation of HepG2, which had been transfected with AdhTIMP-1, was significantly inhibited. Conclusion The proliferation of HepG2 was markedly inhibited by recombinant adenovirus-mediated overexpression of hTIMP-1, which may pave the way for further application in liver gene therapy.