OBJECTIVETo seek possible effect targets of bufalin in HeLa cells by studying the impact of bufalin on cell protein expression profile after treatment on human cervical carcinoma cell lines HeLa.

METHODBufalin's ICs0was measured by MTr assay. The apoptosis of cells was observed by FCM (flow cytometry) and Hoechst 33342 staining assay. Differentiated expression protein spots were founded and identified using proteomic techniques, which could induce HeLa cell apoptosis.

RESULTBufalin showed remarkable cytotoxic effect on HeLa cells. IC50 (154 +/- 21.5) nmol X L(-1) indicated the possibility of inducing cell apoptosis. The protein expression profile showed 11 differentiated expression protein spots. Among the 11 proteins, nudix-type motif 5, vimentin, hnRNP C1/hnRNP C2 variant, HNRPK, HNRPK isoform a variant (two spots are the same protein), heat shock protein 27, macrophage-capping protein, SELENBP1 protein were down-regulated, while ribosomal protein, large, P0 and S-adenosylmethionine synthetase 2 were up-regulated by bufalin treatment. They may be effect targets of bufalin in HeLa cells. Western blotting showed consistent results in heat shock protein 27, vimentin and HNRPK between expression after treatment with bufalin and two-dimensional electrophoresis.

CONCLUSIONBufa-Lin can induce apoptosis in human cervical carcinoma cells HeLa and the effect of bufalin may be related to the joint intervention with multiple protein targets.