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目的:该文通过考察蟾毒灵(bufalin)处理人宫颈癌HeLa细胞株后对细胞蛋白质表达谱的影响,寻找蟾毒灵在HeLa细胞中的可能作用靶点。方法:通过MTT检测细胞活性确定蟾毒灵的IC50,并通过流式细胞术和Hoechst 33342染色检测蟾毒灵诱导细胞凋亡。应用蛋白质组学技术寻找差异表达蛋白点并进行鉴定,用Western blotting试验进行确认。结果:蟾毒灵具有较强的细胞毒作用,IC50(154±21.5)nmol.L-1,可以诱导HeLa细胞发生凋亡。蛋白组学的结果显示蟾毒灵处理组与对照组相比有11个差异表达蛋白,其中9个蛋白点发生了下调;2个蛋白发生了上调。这些蛋白可能是蟾毒灵在HeLa细胞中的作用靶点。Western blotting分析显示heat shock protein 27,vimentin,HNRPK在蟾毒灵处理后的表达调节结果与双向电泳的结果吻合。结论:蟾毒灵可以诱导人宫颈癌HeLa细胞发生凋亡,其凋亡诱导作用可能是多个靶点蛋白共同参与的结果。
OBJECTIVE: To investigate the effect of bufalin on the expression of cell protein in human HeLa cell lines and to find out the possible targets of bufalin in HeLa cells. Methods: The IC50 of bufalin was determined by measuring cell viability by MTT. Bufalin-induced apoptosis was detected by flow cytometry and Hoechst 33342 staining. Proteomics technology was used to find differentially expressed protein spots and identified by Western blotting assay. Results: Bufalin had a strong cytotoxicity. IC50 (154 ± 21.5) nmol.L-1 could induce HeLa cell apoptosis. Proteomics results showed that compared with the control group, there were 11 differentially expressed proteins in the group treated with bufalin, of which 9 were down-regulated and 2 were up-regulated. These proteins may be the target of bufalin in HeLa cells. Western blotting analysis showed that the heat shock protein 27, vimentin, HNRPK expression after bufalin treatment and the two-dimensional electrophoresis results. Conclusion: Bufalin can induce apoptosis in human cervical cancer HeLa cells. The induction of apoptosis may be the result of multiple target proteins.