Experimental Study On Anti-hepatocarcinoma Effect Of Bufalin, An Active Constituent From Arenobufagin

MTT assay and flow cytometry were used to determine the growth inhibition and cell cycle intervention effects of bufalin, cinibufagin and resibufogenin on hepatoma cell lines. Cell apoptosis were detected by FCM, TUNEL labeling, agarose gel electrophoresis, transmission electron microscope. Expression of Bcl-2 and Bax at the protein and mRNA level were detected by immunocytochemical method(ICC) and RT-PCR. Activity of NF- k B and p38MAPK was determined by EMSA and Western blot. Orthotopic transplantation tumour model of human HCC in nude mice was established to be used for the evaluation of anti-tumour effect of bufalin. The reversal effect of bufalin on MDR of BEL-7402/5-Fu cell line was determined by MTT. ADM, Rh-123 and FLU intake in resistant cells was estimated by FCM. Expressions of TS, MRPl, Bcl-xL and Bax proteins were detected by ICC. Bcl-xL and Bax at mRNA level was determined by FQ-PCR. Changes of mitochondrial transmembrane potential( ΔΨm) were determined by FCM. This study come to the conclusion that bufalin has conspicuous anti-hepatocarcinoma activity. Bufalin could induce cell apoptosis by activating NF- κB and p38MAPK passway, and reverse MDR by down-regulating TS and MRPl proteins and regulation of Bcl-xL/Bax ratio to promote the depolarization of ΔΨm.