Combined Inhibitory Effects Of Celecoxib And Fluvastatin On The Growth Of Human Hepatocellar Carcinoma Xenografts In Nude Mice

OBJECTS In this study, we investigated the inhibitory effect of celecoxib, a cyclooxygenase-2 inhibitor, and fluvastatin, a 3-hydroxy-3-methyl glutaryl-CoA reductase inhibitor, on hepatocellar carcinoma in vivo experiments using human hepatocellar carcinoma cell lines, and we aimed to elucidate the possible molecular mechanism of their anticancer effect.METHODS Athymic nude mice implanted with BEL-7402 cells were given celecoxib and fluvastatin, alone and in combination, and their effect on tumor growth was evaluated. Six weeks later, the mice were sacrificed and their tumors were excised for further analysis, Tumor cell proliferation was analyzed by Ki-67 immunostaining. Apoptosis was determined by TUNEL assay. Tumor angiogenesis was investigated by using a monoclonal anti-CD34 Antibody. intratumoral VEGF expression was examined by immunostaining using anti-VEGF monoclonal antibody. Akt, p-Akt, p21waf1/CIP1, mcl-1, and survivin protein levels were measured by Western blot.RESULTS In vivo, combination treatment with celecoxib and fluvastatin resulted in enhanced (a) tumor growth inhibition, (b) apoptosis induction, (c) inhibition of tumor cell proliferation and (d) inhibition of tumor angiogenesis compared with results of single-agent treatment. More over, the combination also enhanced the cyclin-dependent kinase inhibitor p21 waf1/CIP1 expression, decreased the expression of p-Akt, mcl-1 and survivin, but had no effect on the expression of Akt in tumor tissues.CONCLUSION These results suggest that the combination of celecoxib and fluvastatin would be more efficacious for the treatment of HCC than either treatment alone. The present observation may be important in future exploration of the potential application of the combined approach in the treatment of HCC.