Activation,Isolation,Culture And Identification Of Rat Oval Cells And The Role Of P28～(GANK) In Oval Cells Activation And Proliferation

In the current study we activated the rat oval cells by 2-AAF/PH model. The oval cells were dissociated, purified and cultured. The cell immunofluorescence show that these cells immunoreactive to CK19, OV6, albumin, AFP, CD34, CD45, c-kit and Thy1. The rat oval cells can be expanded in vitro culture and induced to differentiate into hepatocyte or cholangiocyte. We also examined the gankyrin protein expression by Western blotting and immunohistochemistry during rat oval cells activation. Gankyrin protein expression was increased in 2-AFF/PH rats in hepatocyte before hepatectomy and decreased after hepatomy at first, and increased expression was found in the activated oval cells. After EGF stimulation, Gankyrin protein was up-regulated in oval cells at 24 to 72 hours, which coincided with increased expression of CyclinD1, CDK4 and decrease of Rb protein. This indicated that p28 plays an important role in oval cells proliferation. We examined in HuH7 stabh transfected with SIRPal derivatives include wild type SIRPal and the tyrosine mutated SIRPal after HGF stimulation. SIRPal inhibited the signal of HGF stimulation and confirmed that SIRPal plays a negative role in hepatocellular carcinoma...