大鼠肝卵圆细胞活化、分离、培养和鉴定及P28～（GANK）在肝卵圆细胞活化和增殖中作用研究

发布时间：2018-12-07 10:05

【摘要】：本研究通过2—AAF/PH模型活化大鼠的肝卵圆细胞,并分离纯化和培养,该细胞表达肝细胞标志如白蛋白、AFP,胆管细胞标志如CK19、OV6,造血干细胞标志c—kit、CD34、CD45、Thy—1。并可以在体外诱导其向肝细胞和胆管细胞分化。同时我们通过Western blot检测发现2—AAF处理后p28~(GANK)表达即升高,于肝切除后先逐渐下降,后在肝卵圆细胞活化过程中又再升高。通过免疫组化发现,肝切除早期,p28~(GANK)主要表达于肝细胞,肝卵圆细胞活化后可见肝卵圆细胞的表达明显高于周围肝细胞。培养的肝卵圆细胞在EGF刺激下,p28~(GANK)表达明显升高,Rb表达下降,CDK4和CycinD1升高,提示p28~(GANK)对肝卵圆细胞活化增殖有重要作用。我们通过HGF刺激稳定表达野生型和突变型SIRPα1(SIRPα1-4Y)以及pcDNA3.0的HuH7细胞进行验证,发现SIRPα1对HGF信号抑制作用,证明了SIRPα1在肝细胞癌的负调控作用。
[Abstract]:In this study, rat hepatic oval cells were activated by 2-AAF/PH model, purified and cultured. The cells expressed hepatocyte markers such as albumin, AFP, bile duct cell markers such as CK19,OV6, hematopoietic stem cell marker c-kitan CD34 and CD45. Thy-1. It can be induced to differentiate into hepatocytes and bile duct cells in vitro. At the same time, we found that the expression of p28 ~ (GANK) increased after 2-AAF treatment, decreased gradually after hepatectomy, and then increased during the activation of hepatic oval cells. It was found by immunohistochemistry that p28 ~ (GANK) was mainly expressed in hepatocytes at the early stage of hepatectomy, and the expression of p28 ~ (GANK) in hepatic oval cells was significantly higher than that in peripheral hepatocytes after activation of hepatic oval cells. Under the stimulation of EGF, the expression of p28 ~ (GANK) increased significantly, the expression of Rb decreased, and the expression of CDK4 and CycinD1 increased, suggesting that p28 ~ (GANK) plays an important role in the activation and proliferation of hepatic oval cells. We tested the expression of wild-type and mutant SIRP 伪 1 (SIRP 伪 1-4Y) and HuH7 cells of pcDNA3.0 by HGF stimulation. We found that SIRP 伪 1 inhibited HGF signal, which demonstrated the negative regulation of SIRP 伪 1 in hepatocellular carcinoma.
【学位授予单位】：第二军医大学
【学位级别】：博士
【学位授予年份】：2005
【分类号】：R329.2

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