肝细胞生长因子和表皮生长因子联合体外诱导大鼠肝卵圆细胞分化的研究

目的探求卵圆细胞向成熟肝细胞分化的演变规律,并对其分化调控的分子机制作初步探讨。方法联合应用肝细胞生长因子和表皮生长因子体外培养大鼠肝脏卵圆细胞OC3,运用电镜、免疫细胞化学、半定量逆转录-聚合酶链反应（RT—PCR）等技术,检测OC3在分化过程中形态及相关分子标记物的改变,并运用蛋白芯片技术观察细胞蛋白表达谱的变化。结果诱导10周后,卵圆细胞胞质变丰富,细胞器增多,表达谷胱甘肽-S-转移酶（GST—P）和丙酮酸激酶（M2-PK）减弱,而表达白蛋白（ALB）、CK18增强,同时诱导后期的细胞较之诱导前主要出现了8种表达差异蛋白。结论在肝细胞生长因子与表皮生长因子的共同作用下,体外培养的卵圆细胞可逐步分化为成熟肝细胞。8种主要的结构蛋白或功能蛋白可能参与了对该过程的调控。

Induced differentiation of rat hepatic oval cells in-vitro by combined hepatocyte growth factor and epidermal growth factor treatment

OBJECTIVE: To characterize the biologic featrues of hepatic oval cells and their protein expression profiles during induced differentiation in vitro. METHODS: Rat hepatic oval cells were treated with epidermal growth factor (EGF) and hepatocyte growth factor (HGF) in vitro, followed by morphological and molecular marker assessment by electromicroscopy, immunocytochemistry, RT-PCR and protein expression chip technology. RESULTS: Ten weeks after induction, the levels of GST-P mRNA and M2-PK mRNA were significantly reduced, whereas those of ALB and CK18 were elevated. Significant variations of expression was seen in 8 protein species during the course of the induced differentiation. CONCLUSION: Combined EGF and HGF treatment in vitro induces cell differentiation of hepatic oval cells, a process in which 8 protein species may play some regulatory roles.