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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 investigate the inhibitory effect of toosendanin on the growth of human triple negative breast cancer MDA-MB-231 cells and its mechanism. Methods MDA-MB-231 cells in logarithmic growth phase were treated with toosendanin at 0, 6.25, 12.50, 25.00, 50.00 and 100.00 nmol / L, and the growth curve was drawn. Toosendanin at 0,12.5 and 50nmol / L for 72h, the detection of toosendanin on the proliferation of MDA-MB-231 cells by MTS, apoptosis and cycle of breast cancer cells by flow cytometry, Western blot detection of apoptosis-related protein Caspase-3, PARP and cleaved-PARP. Results Toosendanin inhibited the proliferation of MDA-MB-231 breast cancer cells in a time and dose-dependent manner (P <0.01). The IC50 of toosendanin at 48, 72 and 96 h were 9.32, 16.96 and 122.37 nmol / L, respectively. Toosendanin could induce apoptosis of breast cancer cells in a concentration-dependent manner (P <0.01), and arrested cells in S phase (P <0.01). The apoptosis rates of MDA-MB-231 cells treated with toosendanin at 0,12.50 and 50.00nmol / L for 72h were 8.12%, 20.85% and 67.21% respectively. The cells in S phase accounted for 32.69% and 47.90% And 61.23% respectively. The apoptosis-related proteins Caspase-3 and PARP decreased and cleaved-PARP increased. Conclusion Toosendanin can inhibit the proliferation of human breast cancer cell line MDA-MB-231, which may be related to the induction of apoptosis and the induction of cell S phase arrest.