川楝素对乳腺癌MDA-MB-231细胞增殖的抑制作用

Inhibitory effect of toosendanin on growth of human breast cancer cells

目的探讨川楝素对人三阴性乳腺癌MDA-MB-231细胞生长的抑制作用及其机制。方法取对数生长期的MDA-MB-231细胞,采用0、6.25、12.50、25.00、50.00和100.00nmol/L川楝素处理,绘制生长曲线。川楝素0、12.5和50nmol/L处理72h后,采用MTS法检测川楝素对MDA-MB-231细胞增殖,流式细胞仪检测乳腺癌细胞凋亡和周期,Western blot检测凋亡相关蛋白半胱氨酶蛋白酶3(Caspase-3)、聚腺苷酸二磷酸核糖转移酶(PARP)和cleaved-PARP表达。结果川楝素呈时间和剂量依赖性抑制MDA-MB-231乳腺癌细胞增殖(P<0.01)。川楝素作用48、72和96h的半数抑制浓度(IC50)分别为9.32、16.96和122.37nmol/L。川楝素能呈浓度依赖性诱导乳腺癌细胞凋亡(P<0.01),阻滞细胞在S期(P<0.01)。以0、12.50和50.00nmol/L川楝素处理MDA-MB-231细胞72h,其早期凋亡率分别为8.12%、20.85%和67.21%,处于细胞周期S期的细胞占32.69%、47.90%和61.23%,凋亡相关蛋白Caspase-3和PARP减少,cleaved-PARP增多。结论川楝素对人乳腺癌MDA-MB-231细胞增殖具有抑制作用;其机制可能与诱导细胞凋亡和引起细胞S期阻滞有关。

Objective To explore the inhibitory effect of toosendanin on the growth of human breast cancer cell line MDA-MB-231 cells and its possible mechanism. Methods The MDA-MB-231 cell growth curve was drawn by treating the cells in logarithmic growth phase with different concentrations of toosendanin 0,6.25,12. 50,25.00,50.00 and 100. 00 nmol/L. After treating the cells with toosendanin 0,12.5 and 50 umol/L for 72 hours, the effect of toosendanin on the proliferation of MDA-MB-231 cells was observed with MTS assay, cell apoptosis and cycle were measured by flow cytometry, and the expressions of Caspase-3, PARP and cleaved-PARP were detected by Western blot. Results The proliferation of MDA-MB-231 cells was significantly inhibited by toosendanin in the dose- and time-dependent manners （P〈0. 01 ）. The half inhibitory cencentrations （IC50） were 9.32 nmol/L,16. 96 nmol/L and 122. 37 nmol/L for the treating times of 48 hours,72 hours and 96 hours, respectively. Toosendanin inhibited the apoptosis of MDA-MB-231 cells in a concentration-dependent manner（P〈0.01） and blocked them in S phase. The expressions of Caspase-3 and PARP were decreased, but cleaved-PARP expression was increased. Conclusion Toosendanin can inhibit the growth of breast cancer cells, which is possibly related to inducing cell apoptosis and bloching cell cycle in S-phase.