川楝素对多柔比星体外抗乳腺癌活性的影响

Toosendanin Enhances the Anti-tumor Effect of Doxorubicin on Breast Cancer in vitro by Upregulating the Expression of FOXO1

目的观察川楝素对多柔比星体外抗乳腺癌活性并研究其机制。方法多柔比星(0~3μg/m L)和川楝素(10mol/L)处理细胞48h,MTT法检测乳腺癌细胞系MDA-MB-435和MDA-MB-231细胞活力。转染后的MDA-MB-435细胞经多柔比星(0.2μg/m L)和川楝素(10mol/L)处理48h,流式细胞术检测MDA-MB-435细胞凋亡;Western blot实验检测MDA-MB-435细胞FOXO1、Bim、Noxa表达水平,细胞色素C的释放水平及半胱氨酸蛋白酶(Caspase)-9、Caspase-3活化水平。结果川楝素明显增强MDA-MB-435和MDA-MB-231细胞系对不同浓度多柔比星的敏感性。多柔比星联合川楝素处理的MDA-MB-435相对细胞活力(0.55±0.04)显著低于多柔比星单治疗组(0.87±0.07,P<0.05)和多柔比星+川楝素+FOXO1 siRNA组(0.79±0.07,P<0.05)。多柔比星联合川楝素处理的MDA-MB-435细胞凋亡率[(34.2±2.8)%]显著高于多柔比星单治疗组[(9.3±0.8)%,P<0.05]和多柔比星+川楝素+FOXO1 siRNA组[(12.5±1.1)%,P<0.05]。多柔比星联合川楝素处理的MDAMB-435细胞中FOXO1、Bim、Noxa表达水平,细胞色素C的释放水平及Caspase-9、Caspase-3活化水平均显著高于多柔比星单治疗组和多柔比星+川楝素+FOXO1 siRNA组。结论川楝素可能通过上调FOXO1表达增强多柔比星的体外抗乳腺癌活性。

Objective To investigate the adjuvant effect and mechanism of toosendanin on enhancing the anti- tumor effect of doxorubicin against breast cancer. Methods MTr assay was performed to measure the cell viability of MDA-MB-435 and MDA-MB-231 after 48h cocuhured with doxorubicin （0-3μg/mL） and toosendanin（10mol/L）. Flow cytometry analysis was performed to detect the cell apoptosis of MDA-MB-435 treated with doxorubicin （0.2μg/mL） and toosendanin （lOmol/L） for 48h; western blot analysis was performed to evaluate the expression of FOXO1, Bim, Noxa, and release of cytochrome c, and the activation of caspase-9 and caspase-3 in doxorubicin （0.2μg/mL） and toosendanin （10mol/L）-treated MDA-MB-435 cells. Results Toosendanin significantly increased the sensitivity of MDA-MB-435 and MDA-MB-231 cell lines to different concentrations of doxornbicin. Relative cell viability of MDA-MB-435 co-treated with doxornbicin and toosendaninwas significantly （0.55±0.04） lower than that in the doxorubicin single treatment group[（ 0.87±0.07 ）, P〈0.05] and the doxorubicin±toosendanin±FOXO1 siRNA group （0.79±0.07, P〈0.05）. Apoptotic rate of MDA-MB-435 co-treated with doxornbicin and toosendanin （34.2%± 2.8%） was significantly higher than that in the doxornbicin single treatment group （9.3%_±0.8%, P〈0.05） and dox- orubicin±toosendanin±FOXO1 siRNA group（ 12.5%±1.1%, P〈0.05）. Expression of FOXO1, Bim, Noxa, and release of cytochrome c, and activation of caspase-9 and caspase-3 in MDA-MB-435 cells co-treated with doxorubicin and toosendanin was significantly increased than those in the doxorubicin single treatment group and doxornbicin ＋ toosendanin±FOXO1 siRNA group （all P〈0.05）. Conclusion Toosendanin may enhance the anti-tumor effect of doxornbicin on breast cancer in vitro by upregulating the expression of FOXO1.