薯蓣皂苷元对盐酸米托蒽醌诱导耐药子宫肉瘤细胞MES-SA/MX2凋亡的增敏作用

Sensitizing Effect of Diosgenin on the Apoptosis Induced by Mitoxantrone Hydrochloride in MDR Uterine Sarcoma Cell Lines( MES-SA/MX2)

目的研究薯蓣皂苷元对盐酸米托蒽醌的抗肿瘤增敏作用及其机制,为临床联合用药提供指导。方法以维拉帕米为阳性对照,利用四甲基偶氮唑蓝(MTT)法测定薯蓣皂苷元和抗癌药盐酸米托蒽醌联用后对耐药细胞株MES-SA/MX2增殖的影响;蛋白印迹法(WB)和实时荧光定量PCR(RT-q PCR)法研究薯蓣皂苷元对细胞P-糖蛋白(P-gp)的表达影响,流式细胞仪测定候选化合物对P-gp底物四甲基罗丹明甲酯(TMRM)在细胞中的摄取,试剂盒测定ATP酶活性。结果薯蓣皂苷元可增加盐酸米托蒽醌对耐药细胞株MES-SA/MX2的增殖抑制,逆转指数为维拉帕米的2.4倍,同时降低P-gp蛋白达22.2%(P<0.01)的表达,但对P-gp的基因表达无影响,能提高TMRM的胞内荧光强度达14.5%,且不影响ATP酶活性。结论薯蓣皂苷元增强米托蒽醌对耐药子宫瘤细胞的敏感性,其作用与抑制P-gp功能有关。

OBJECTIVE To investigate the sensitizing effect and mechanism of diosgenin on anti-proliferation activity of mitox- antrone hydrochloride on MDR cells. METHODS Verapamil was used as positive control. MTI＂ method was used to evaluate the pro- liferation inhibition effect of diosgenin combined with mitoxantrone hydrochloride on MES-SA/MX2 cells. Western blot method （WB） and RT-qPCR were used to evaluate the effect of diosgeninon protein and gene expression of P-gp respectively. The uptake activity of TMRM was measured by flow cytometry（FCM）. ATPase activity was determined by kit. RESULTS Diosgenin can ehhance prolifera- tion inhibition of mitoxantrone hydrochloride in MDR cell lines （MES-SA/MX2）. The reversing index was 2.4 times of verapamil. Di- osgenin decreased P-gp protein expression by 22. 2% （ P 〈 0. 01 ） , but had marginal effect on P-gp mRNA. Diosgenin increased the fluorescence intensity of TMRM by 14. 5% , and did not affect ATPase activity. CONCLUSION Diosgenin can increase the sensitiv- ity of mitoxantrone hydrochloride by inhibiting P-gp.