江苏大学学报（医学版）2024，Vol.34Issue(4)：324-330,7.DOI:10.13312/j.issn.1671-7783.y230123
姜黄素通过降低miR-135b-5p和FEN1表达增强卵巢癌OVCAR-3细胞顺铂敏感性
Curcumin enhances cisplatin sensitivity of OVCAR-3 cells by decreasing the expression of miR-135b-5p and FEN1
摘要
Abstract
Objective:To investigate the effect of curcumin on cisplatin(DDP)sensitivity of ovarian cancer OVCAR-3 cells and its potential mechanism.Methods:(1)Ovarian cancer parental cell line OVCAR-3 was routinely cultured in vitro and cisplatin-resistant cell line OVCAR-3/DDP was constructed.OVCAR-3 cells were divided into Control(conventional culture),DDP-1(1 μmol/L cisplatin),DDP-5(5 μmol/L cisplatin),DDP-10(10 μmol/L cisplatin),DDP-15(15 μmol/L cisplatin),DDP-20(20 μmol/L cisplatin),DDP-30(30 μmol/L cisplatin),curcumin+DDP-1(20 μmol/L curcumin+1 μmol/L cisplatin),curcumin+DDP-5(20 μmol/L curcumin+5 μmol/L cisplatin),curcumin+DDP-10(20 μmol/L curcumin+10 μmol/L cisplatin),curcumin+DDP-15(20 μmol/L curcumin+15 μmol/L cisplatin),curcumin+DDP-20(20 μmol/L curcumin+20 μmol/L cisplatin)and curcumin+DDP-30 groups(20 μmol/L curcumin+30 μmol/L cisplatin).OVCAR-3/DDP cells were divided into Control,DDP-10(10 μmol/L cisplatin),DDP-20(20 μmol/L cisplatin),DDP-30(30 μmol/L cisplatin),DDP-40(40 μmol/L cisplatin),DDP-50(50 μmol/L cisplatin),DDP-60(60 μmol/L cisplatin),curcumin+DDP-10(20 μmol/L curcumin+10 μmol/L cisplatin),curcumin+DDP-20(20 μmol/L curcumin+20 μmol/L cisplatin),curcumin+DDP-30(20 μmol/L curcumin+30 μmol/L cisplatin),curcumin+DDP-40(20 μmol/L curcumin+40 μmol/L cisplatin),curcumin+DDP-50(20 μmol/L curcumin+50 μmol/L cisplatin),curcumin+DDP-60(20 μmol/L curcumin+60 μmol/L cisplatin)groups.CCK-8 method was used to detect the cell viability,the median inhibitory concentration of cisplatin(IC50)was calculated,and the appropriate concentration of cisplatin and curcumin was screened.(2)OVCAR-3 cells were divided into Control group,DDP group(20.5 μg/mL cisplatin),curcumin group(20 μmol/L curcumin)and curcumin+DDP group(20 μmol/L curcumin+20.5 μg/mL cisplatin).OVCAR-3/DDP cells were divided into Control group,DDP group(42.1 μg/mL cisplatin),curcumin group(20 μmol/L curcumin)and curcumin+DDP group(20 μmol/L curcumin+42.1 μg/mL cisplatin).The apoptosis rate was detected by flow cytometry,the expression of miR-135b-5p was detected by quantitative real-time PCR(qRT-PCR),and the mRNA and protein expressions of flap endonuclease 1(FEN1)were detected by qRT-PCR and Western blotting,respectively.Results:Compared with DDP group,the activity of OVCAR-3 and OVCAR-3/DDP cells in curcumin+DDP group was significantly decreased at the same cisplatin concentration(P＜0.001).The IC50 of DDP on OVCAR-3 and OVCAR-3/DDP cells were 20.5 μg/mL and 42.1 μg/mL,respectively.Compared with OVCAR group or OVCAR/DDP group,curcumin or cisplatin significantly increased the apoptosis rate of OVCAR-3 and OVCAR-3/DDP cells(all P＜0.001),and the combined effect of curcumin and cisplatin was more significant.In addition,curcumin or cisplatin treatment significantly decreased the expression of miR-135b-5p and FEN1 expression in OVCAR-3 and OVCAR-3/DDP cells(P＜0.01 or＜0.001),and the combined effect was more significant.Conclusion:Curcumin could enhance cisplatin sensitivity in ovarian cancer OVCAR-3 cells,which may be attributed to the reducing expression of miR-135b-5p and FEN1.关键词
姜黄素/miR-135b-5p/瓣状核酸内切酶-1(FEN1)/卵巢癌/顺铂敏感性Key words
Curcumin/miR-135b-5p/flap endonuclease 1(FEN1)/ovarian cancer/cisplatin sensitivity分类
医药卫生引用本文复制引用
马蓉,李娟,王芳..姜黄素通过降低miR-135b-5p和FEN1表达增强卵巢癌OVCAR-3细胞顺铂敏感性[J].江苏大学学报（医学版）,2024,34(4):324-330,7.基金项目
新疆维吾尔自治区自然科学基金资助项目(2021D01C237) （2021D01C237）
