Abstract:

To isolate，cultivate and identify the hepatocellular carcinoma associated fibroblasts (hCAF) and to investigate their possible role in the occurrence and development of hepatocellular carcinoma.Methods The primary hCAF were isolated and purified by enzyme digestion method. The expressions of the specific proteins associated with hCAF were examined by Western blotting method. The changes of cell cycles of Hep3B cells after hCAF supernatant treatment were evaluated by flow cytometry. The effect of hCAF on hepatocellular carcinoma cells  was observed though building a xenograft tumor model in nude mice. Results The hCAF were successfully isolated and confirmed and highly specificly  expressed α-smooth muscle actin (α-SMA)  detected  by Western blotting method. The proliferation capacity of Hep3B was increased significantly after treated with hCAF conditioned media. The proliferation capacities of Hep3B were 126.0%,125.0% and 120.5% of those treated with complete medium for 48 h(P<0.05).The number of carcinoma cells and the percentages of carcinoma cells in S phase were higher than those treated with complete medium for  48 h(P<0.05).The tumor volume in Hep3B cells and hCAF co-injection group ［(1.34±0.52) cm2］ was significantly larger than that in Hep3B cells inoculation  group ［(0.51±0.09)cm2］ in nude mice(P<0.05). Conclusion hCAF could promote the proliferation of hepatocellular carcinoma cell line Hep3B, and  hCAF might play an important role in the progression of hepatocellular carcinoma.

Key words: carcinoma associated fibroblast, hepatocellular tumor, proliferation, tumor microenvironment