重组人肝刺激因子的促增殖和抗损伤作用

INCREASED CELLULAR PROLIFERATION AND PROTECTION OF RECOMBINED HUMAN HEPATIC STIMULATOR SUBSTANCE

目的 探讨重组人肝刺激因子 (hHSS)的生物学特性并观察其抗损伤能力。 方法 以MTT法、细胞计数、细胞周期分析和MAPK活性观察hHSS促细胞增殖作用。以H2 O2 损伤细胞 ,观察hHSS保护细胞的作用。 结果 重组hHSS能刺激BEL 74 0 2肝癌细胞增殖 ,促进增殖信号分子MAPK的磷酸化。hHSS可增强细胞对抗H2 O2损伤的能力 ,促进受损细胞DNA合成。 结论 HSS为一种新的生物活性蛋白 。

Objective Based on the result that a gene coding for human HSS had been identified and prokaryotically expressed, this study is aiming to investigate the proliferative and protective effect of recombinant HSS on hepatoma cells. Methods hHSS protein was produced in BL\|21 strain of E.Coli containing pET\|42a vector and purified with His\5Tag affinity chromatography. A various dosages of 80\|400?μg/L were administrated into cell culture. The cell proliferation was analyzed by MTT, cell\|count and flow cytometry methods. Furthermore, cellular growth signaling as indexed by phosphorylation of mitogen\|activated protein kinase (MAPK) was determined with Western blot. In addition to cell growth, protection of hHSS on the cells against H\-2O\-2 was also observed. Results It was showed that recombinaht HSS of 400?μg/L was able to promote DNA synthesis by 21\^5% as compared to non\|treated cells. After 24?h of HSS action, the cell division measured by MTT method as well as cell\|count was significantly enhanced. Meantime, it was indicated that the phosphorylation of MAPK Thr202/Tyr204 was increased in 79\^0% as compared with that of the non\|treated cells. Pretreatment of the cells with hHSS for 12?h prior to H 2O 2 injury preserved the survival of them.Conclusion It is postulated that hHSS is an active protein to stimulate liver proliferation. [