厚朴酚通过改变Bax/Bcl-X_L表达和激活caspase诱导人宫颈癌HeLa细胞凋亡

Magnolol induces apoptosis via alteration of Bax/Bcl-X_L ratio and activation of caspases

目的研究厚朴酚诱导HeLa细胞凋亡机制。方法MTT法测定厚朴酚对HeLa和人正常胚肺HEL299细胞的细胞毒作用。通过显微镜,荧光染色观察细胞形态学变化,用琼脂糖凝胶电泳检测DNA片断化。用Western印迹法分析药物对蛋白质表达的影响。结果厚朴酚明显抑制HeLa细胞的增殖,诱导HeLa细胞调亡。对HEL299细胞的细胞毒作用弱于HeLa细胞。Caspase3,8,9,10及家族抑制剂可明显抑制厚朴酚诱导的凋亡。激动型Fas抗体CH11对厚朴酚诱导的HeLa细胞死亡有协同作用。Western印迹结果显示厚朴酚作用12h后Bax/BclXL表达比率升高,caspase3前体及其底物ICAD和PARP发生降解,磷酸化p53和p53蛋白表达增加。结论厚朴酚(80μmol·L-1)通过改变Bax/BclXL的表达激活caspase途径诱导HeLa细胞凋亡。

Aim To study the mechanism of magnolol-induced HeLa cell death.Methods Cell viability was measured by MTT method.Morphological changes were observed by phase contrast microscopy and Hoechst 33258 staining. DNA fragmentation was assayed by agarose gel electrophoresis. Protein level was detected by Western blot analysis.Results Magnolol induced HeLa cell apoptosis and had a weaker cytotoxic effect on HEL 299 cell. The apoptosis of HeLa cells was partially reversed by caspase-3,-8,-9,-10 and caspase family inhibitors. Magnolol has a synergistic apoptotic effect with Fas agonistic antibody CH-11. Treatment of HeLa cells with magnolol for 12 h increased the protein expression ratio of Bax/Bcl-X_L; procaspase-3, ICAD and PARP were cleaved to smaller molecules. p53 and phosphorylation of p53 (ser-15) increased response to magnolol treatment.Conclusion Magnolol induced HeLa cell death via alteration of Bax/Bcl-X_L ratio,activation of caspases and accumulation of p53.