Herbal formula of Bushen Jianpi(补肾健脾方)combined with sorafenib inhibits hepatocellular carcinoma growth by promoting cell apoptosis and blocking the cell cycle

OBJECTIVE:To investigate the efficacy of an herbal formula of Bushen Jianpi(补肾健脾方,BSJP)combined with sorafenib on hepatocellular carcinoma(HCC)in vitro and in vivo,and to study the underlying mechanisms of action.METHODS:BSJP,a mixture of 12 raw herbs,was extracted in 70%alcohol/30%water and freeze-dried into a powder.The in vitro effects of BSJP alone,sorafenib alone,and their combination on cell survival,apoptosis,and cell cycle distribution were evaluated in HCC cell lines HCCLM3,HepG2,and SMMC-7721.The expression of B-cell lymphoma-2(Bcl-2),caspase-3,and caspase-9 in HCCLM3 cells was measured using Western blots after drug administration.The in vivo effects of BSJP and sorafenib were evaluated in a tumor surgical resection model using 4-week old male athymic BALB/c nude mice injected with HCCLM3 cells.Immunohistochemical analysis of tumor tissues was performed to evaluate the effects of BSJP alone,sorafenib alone,and their combination on the expression of caspase-3,caspase-9,and Bcl-2.RESULTS:BSJP decreased the survival rate of HCC cell lines,and the combination of BSJP and sorafenib further decreased the survival rate.BSJP significantly promoted cell apoptosis and blocked cell-cycle progression in HCCLM3,HepG2,and SMMC-7721 cells in a dose-dependent manner.Furthermore,the administration of BSJP and sorafenib inhibited the growth of HCCLM3 cell xenografts in nude mice,with no reduction in body weight.In vivo and in vitro experiments showed that BSJP combined with sorafenib could significantly decrease the expression of Bcl-2.CONCLUSION:Our findings suggest that the herbal formula of BSJP is a potential HCC antitumor agent.