五味子乙素促进多柔比星诱导肝癌细胞SMMC7721凋亡的研究

Schisandrin B enhances doxorubicin-induced apoptosis in SMMC7721 cell line

目的验证五味子乙素对多柔比星(阿霉素)诱导肝癌细胞SMM C 7721凋亡的促进作用。方法M TT法测定药物对细胞存活的影响;流式细胞仪P I染色测定药物引起的细胞凋亡率,R-藻红蛋白标记测定P糖蛋白表达,R hodanm in 123(R h123)标记检测线粒体膜电位变化;流式细胞仪测定五味子乙素对阿霉素积聚外排的影响;W estern b lot检测药物作用后相关蛋白(caspase-3和PARP)表达。结果五味子乙素能显著促进阿霉素诱导人肝癌细胞系SMM C 7721凋亡(P<0.01),但不增强阿霉素对正常细胞(大鼠心肌细胞和人纤维原细胞)的毒性作用。这种增敏作用与五味子乙素对P糖蛋白及其它药泵的抑制作用无关,但与caspase的激活相关。结论五味子乙素能促进阿霉素诱导的肝癌细胞凋亡,但不增加阿霉素对正常细胞的毒性作用。

Objective To demonstrate the capability of Schisandrin B （SchB） in enhancing the doxorubicin （DOX）- induced apoptosis in human hepatic carcinoma cell line SMMC7721. Methods The influence of Sch B on cell survival was evaluated by MTT assay. The drug-induced apoptotic rate was evaluated by flow cytometry（FCM） with PI staining. The expression of P-glycoprotein,the loss of mitochondria membrane potential and the effect of Sch B on DOX accumulation and efflux were investigated by FCM after the treatments of 5μmol/L Dox with or without 50μmol/L Sch B for appropriate time. And the expression of related proteins （caspase-3 and PARP） after drug treatments was investigated by Western blotting. Results 50 μmol/L Sch B significantly enhanced 5 μmol/L Dox-induced apoptosis in SMMC7721 cell line （P〈0. 01）. This effect was irrelevant with the inhibition of P-glycoprotein or other drug pump ,but was related to the activation of caspase-3. Conclusion As a sensitizer,the adoption of Sch B can effectively enhance DOX-induced apoptosis in cancer cell lines,but not enhance DOX-induced apoptosis in normal cells.