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目的观察中药成份华蟾毒精(CBG)对HeLa细胞增殖的作用,并初步探讨其作用机理。方法SRB和MTT法观察CBG对人肝癌细胞(Bel-7402)、宫颈癌细胞(HeLa)、乳腺癌细胞(MCF-7)、胃癌细胞(BGC-823)和白血病淋巴细胞(HL60)等几种人体肿瘤细胞增殖的影响;采用流式细胞仪法观察CBG对HeLa细胞周期的影响;通过双向电泳的方法,进一步探讨CBG作用于HeLa细胞蛋白质组的情况。结果CBG对多种人体肿瘤细胞的增殖都有明显抑制作用,其Bel-7402、HeLa、MCF-7、BGC-823和HL60细胞的IC50值分别为0.011,0.019,0.116,0.149,1.369μmol/L,其中以人宫颈癌细胞(HeLa)和人肝癌细胞(Bel-7402)最为敏感。流式方法测定的结果显示,不同浓度的CBG与HeLa细胞作用72h,可使G2/M期的HeLa细胞由17.3%增加到35.6%。双向电泳的结果显示,0.02μmol/L CBG作用48h后,与对照组相比,HeLa细胞中相对分子质量较小(约30000~90000)的偏酸性蛋白(pH约4~6)发生了较大的改变。结论CBG对多种人体肿瘤细胞的增殖均有显著抑制作用。CBG可使HeLa细胞周期阻滞在G2/M期,并对HeLa细胞内相对分子质量较小的酸性蛋白表达有明显的影响。
Objective To observe the effect of Chinese medicinal ingredient canadonin (CBG) on the proliferation of HeLa cells and to explore its mechanism. Methods The effects of CBG on Bel-7402, HeLa, MCF-7, BGC-823 and HL60 cells were observed by SRB and MTT assay. The effect of CBG on HeLa cell cycle was observed by flow cytometry. The effect of CBG on HeLa cell proteome was further explored by two-dimensional electrophoresis. Results The IC50 of Bel-7402, HeLa, MCF-7, BGC-823 and HL60 cells were significantly lower than that of CBG cells (P <0.01) , Of which human cervical cancer cells (HeLa) and human hepatocellular carcinoma cells (Bel-7402) the most sensitive. Flow cytometry results showed that different concentrations of CBG and HeLa cells for 72h, G2 / M phase HeLa cells increased from 17.3% to 35.6%. The results of two-dimensional electrophoresis showed that after treated with 0.02μmol / L CBG for 48h, the relative molecular mass of HeLa cells (about 30,000 ~ 90,000) was larger than that of the control group (pH about 4 ~ 6) Change. Conclusion CBG has a significant inhibitory effect on the proliferation of many human tumor cells. CBG can block HeLa cell cycle in G2 / M phase and have a significant effect on the expression of acidic protein with relatively low molecular weight in HeLa cells.