芍药内酯苷对人卵巢癌多药耐药的逆转作用

The reversal effect of albiflorin on multidrug resistance of human ovarian cancer and its mechanism

目的研究芍药内酯苷(albiflorin,AL)逆转人卵巢癌多药耐药的作用和潜在机制。方法采用CCK-8试剂盒检测AL对SKOV3/DDP细胞的耐药逆转作用,采用流式细胞术检测AL对P糖蛋白(P-glycoprotein,P-gp)功能的影响,采用RT-qPCR和Western blot检测AL对SKOV3/DDP细胞中MYC、WWP1和ABCB1/P-gp的影响。采用RNA干扰技术构建MYC敲低的SKOV3/DDP细胞株,观察其耐药性、P-gp功能及相关基因和蛋白表达变化。结果AL对SKOV3/DDP细胞有耐药逆转作用,对P-gp功能抑制作用具有浓度依赖性。AL 25、50、100μmol·L^(-1)对ABCB1/P-gp、MYC和WWP1的抑制作用均逐渐增强。MYC抑制剂MYCi975对ABCB1/P-gp、MYC和WWP1的抑制作用均强于或相当于AL 100μmol·L^(-1)组。SKOV3/DDP细胞敲低MYC后,细胞耐药性、P-gp功能以及相关基因和蛋白表达均受到抑制。结论AL对SKOV3/DDP细胞的耐药逆转作用可能与抑制P-gp功能和ABCB1/P-gp、MYC、WWP1表达有关,为AL作为耐药逆转剂用于卵巢癌的临床治疗提供了实验基础。

Aim To study the reversal effect of albiflorin(AL)on multidrug resistance of human ovarian cancer and the potential mechanism.Methods The drug resistance reversal effect of AL on SKOV3/DDP cells was detected by CCK-8 kit,and the effect of AL on P-glycoprotein(P-gp)function was detected by flow cytometry.The effects of AL on MYC,WWP1 and ABCB1 in SKOV3/DDP cells were detected by RT-qPCR and Western blot.The MYC-knockdown SKOV3/DDP cell line was constructed by RNA interference technology,and its drug resistance,P-gp function and related gene and protein expression changes were investigated.Results AL had a drug resistance reversal effect on SKOV3/DDP cells and a concentration-dependent inhibitory effect on P-gp function.The inhibitory effects of AL 25,50 and 100μmol·L^(-1) on ABCB1/P-gp,MYC and WWP1 were gradually enhanced.The inhibitory effect of MYCi975,a MYC inhibitor,on ABCB1/P-gp,MYC and WWP1 was stronger than or equivalent to that of AL 100μmol·L^(-1) group.After knockdown of MYC in SKOV3/DDP cells,cell drug resistance,P-gp function,and related gene and protein expression were inhibited.Conclusions The drug resistance reversal effect of AL on SKOV3/DDP cells may be related to the inhibition of P-gp function and the expression of ABCB1/P-gp,MYC and WWP1,which provides an experiment base for the development of AL as a drug resistance reversal agent for the clinical treatment of ovarian cancer.