牛蒡子苷元对卵巢癌细胞耐药性的影响

Effect of arctigenin on drug resistance in ovarian cancer cells

目的:探讨牛蒡子苷元对人卵巢癌顺铂耐药细胞株SKOV3/DDP细胞耐药性的影响及其可能的作用机制。方法:构建卵巢癌耐药细胞株SKOV3/DDP。用不同浓度的牛蒡子苷元(1、5、10、15、20、25 mmol/L)分别处理SKOV3和SKOV3/DDP细胞,CCK-8法检测细胞生长,筛选适用浓度。随后用10 mmol/L牛蒡子苷元处理SKOV3/DDP细胞。流式细胞仪检测细胞凋亡,Western blot检测Caspase-3/9和Cleaved Caspase-3/9蛋白表达。CCK-8法检测细胞半数抑制浓度(50%inhibition concentration,IC_(50))。RT-PCR和Western blot分别检测Survivin的mRNA和蛋白表达。在牛蒡子苷元处理的SKOV3/DDP细胞中转染Survivin过表达质粒检测IC_(50)和细胞凋亡水平。结果:不同浓度牛蒡子苷元对SKOV3和SKOV3/DDP细胞生长均有抑制作用,并且当牛蒡子苷元的浓度≥10 mmol/L时,对SKOV3/DDP细胞生长抑制作用高于SKOV3细胞(P<0.05)。10 mmol/L牛蒡子苷元的处理显著提升SKOV3/DDP细胞凋亡,增加凋亡蛋白Caspase-3/9和Cleaved Caspase-3/9的表达,降低细胞顺铂IC_(50)值,并抑制细胞中Survivin的mRNA和蛋白表达(P<0.05)。在转染Survivin过表达质粒后,牛蒡子苷元处理的SKOV3/DDP细胞IC_(50)值显著提升,细胞凋亡率显著下降(P<0.05)。结论:牛蒡子苷元可能通过抑制Survivin降低卵巢癌耐药细胞SKOV3/DDP的耐药性。

Objective:To explore the effect and underlying mechanism of arctigenin on drug resistance of human ovarian cancer cisplatin resistant cell line SKOV3/DDP.Methods:Establish ovarian cancer drug resistant cell line SKOV3/DDP.SKOV3 and SKOV3/DDP were treated with arctigenin at different concentration(1,5,10,15,20,25 mmol/L).CCK-8 method was used to measure cell proliferation and screen optimum drug concentration.Then SKOV3/DDP were treated with 10 mmol/L arctigenin.Cell apoptosis was measured by flow cytometry.Protein expression of Caspase-3/9 and Cleaved Caspase-3/9 were measured by Western blot.IC_(50)were measured by CCK-8 method.mRNA and protein expression of Survivin were measured by RT-PCR and Western blot,respectively.SKOV3/DDP cell were treated with arctigenin and transfected with Survivin overexpression plasmid,then IC_(50)and cell apopotsis were texted.Results:Different concentrations of arctigenin inhibited cell growth of both SKOV3 and SKOV3/DDP.The inhibitory effect of arctigenin was higher in SKOV3/DDP than SKOV3 cells when the concentration of arctigenin ≥10 mmol/L(P<0.05).10 mmol/L arctigenin treatment significantly increased SKOV3/DDP cell apoptosis,elevated protein expression of Caspase-3/9 and Cleaved Caspase-3/9,decreased DDP IC_(50),and inhibited mRNA and protein expression of Survivin(P<0.05).After transfection of Survivin overexpression vector,IC_(50)was elevated and cell apoptosis was decreased in arctigenin treated SKOV3/DDP cells(P<0.05).Conclusion:Arctigenin might inhibited drug resistance of SKOV3/DDP cells through suppressing Survivin.