芒柄花素抑制自噬逆转激素受体阳性人乳腺癌细胞多药耐药的机制研究

Mechanism of formononetin on reversing multidrug resistance in hormone receptor-positive human breast cancer cells via autophagy inhibition

目的观察芒柄花素对激素受体阳性人乳腺癌阿霉素耐药细胞MCF-7/ADR的多药耐药逆转作用及其对自噬相关蛋白LC3A/B、Beclin-1、VPS34和耐药相关蛋白P-gp的影响。方法芒柄花素、阿霉素(多柔比星Doxorubicin,DOX)、紫杉醇(Paclitaxel,PTX)、长春瑞滨(Vinorelbine,NVB)处理MCF-7及MCF-7/ADR细胞,MTT法检测芒柄花素、DOX、PTX、NVB对MCF-7及MCF-7/ADR细胞毒性的影响,以及不同浓度的芒柄花素对上述化疗药物作用下MCF-7及MCF-7/ADR细胞毒性的影响,Western blot法检测芒柄花素作用于MCF-7及MCF-7/ADR后耐药及自噬相关蛋白的变化。结果①MCF-7/ADR对多种化疗药物均耐药;②芒柄花素对肿瘤细胞生长抑制作用很弱,且有抗肿瘤耐药活性;③在耐药细胞MCF-7/ADR细胞中,使用不同浓度的芒柄花素(1、3、10、30、100μmol/L)与这些抗肿瘤药物合用后,IC50值与不用芒柄花素组相比,差异有统计学意义(P<0.05);④随着芒柄花素浓度的增加,耐药蛋白P-gp的表达呈下降趋势,自噬标志物LC3A/B呈下降趋势,VPS-34、Beclin-1蛋白变化不明显。结论芒柄花素能逆转激素受体阳性人乳腺癌阿霉素耐药细胞MCF-7/ADR多药耐药,其作用机制可能与芒柄花素能抑制自噬发生、降低P-gp蛋白有关。

Objective To observe the reversal effects of formononetin on multidrug resistance in hormone receptor positive human breast cancer adriamycin-resistant cells MCF-7/ADR,and its effects on autophagy-related proteins LC3 A/B,Beclin-1,VPS34 and resistance-related protein P-gp.Methods MCF-7 and MCF-7/ADR cells were treated with formononetin,doxorubicin(DOX),paclitaxel(PTX)and vinorelbine(NVB).The effects of formononetin,DOX,PTX,NVB on the cytotoxicity of MCF-7 and MCF-7/ADR cells were detected by MTT assay,as well as the effects of formononetin at different concentrations on the cytotoxicity of MCF-7 and MCF-7/ADR cells induced by above chemotherapeutics.After treatment with formononetin,the changes of drug resistance and autophagy related proteins in MCF-7 and MCF-7/ADR cells were detected by Western blot.Results①MCF-7/ADR cells was resistant to many kinds of chemotherapeutic drugs.②Formononetin showed weak inhibitory effect on tumor cell growth,but inhibited the tumor drug resistance.③In drug-resistant MCF-7/ADR cells,formononetin at different concentrations(1,3,10,30,100μmol/L)were added combined with these chemotherapeutic drugs,there was statistically significant difference in IC50 value compared with the groups without formononetin intervention(P<0.05).④With the increase of formononetin concentration,the expression of drug-resistant protein P-gp showed a downward trend,and the autophagy marker LC3 A/B also showed a downward trend,but no obvious changes on the VPS-34 and Beclin-1 proteins were found.Conclusion Formononetin can reverse multidrug resistance of hormone receptor positive human breast cancer adriamycin-resistant cells MCF-7/ADR.Its mechanism maybe formononetin can inhibit autophagy and reduce P-gp protein.