摘要
目的:探究姜黄素对卵巢癌OVCAR-3细胞顺铂敏感性的影响及其潜在机制。方法:(1)体外常规培养卵巢癌亲本细胞OVCAR-3并构建顺铂耐药OVCAR-3/DDP细胞,将OVCAR-3细胞分为Control(常规培养)、DDP-1(1μmol/L顺铂)、DDP-5(5μmol/L顺铂)、DDP-10(10μmol/L顺铂)、DDP-15(15μmol/L顺铂)、DDP-20(20μmol/L顺铂)、DDP-30(30μmol/L顺铂)、姜黄素+DDP-1(20μmol/L姜黄素+1μmol/L顺铂)、姜黄素+DDP-5(20μmol/L姜黄素+5μmol/L顺铂)、姜黄素+DDP-10(20μmol/L姜黄素+10μmol/L顺铂)、姜黄素+DDP-15(20μmol/L姜黄素+15μmol/L顺铂)、姜黄素+DDP-20(20μmol/L姜黄素+20μmol/L顺铂)、姜黄素+DDP-30组(20μmol/L姜黄素+30μmol/L顺铂);将OVCAR-3/DDP细胞分为Control(常规处理)、DDP-10(10μmol/L顺铂)、DDP-20(20μmol/L顺铂)、DDP-30(30μmol/L顺铂)、DDP-40(40μmol/L顺铂)、DDP-50(50μmol/L顺铂)、DDP-60(60μmol/L顺铂)、姜黄素+DDP-10(20μmol/L姜黄素+10μmol/L顺铂)、姜黄素+DDP-20(20μmol/L姜黄素+20μmol/L顺铂)、姜黄素+DDP-30(20μmol/L姜黄素+30μmol/L顺铂)、姜黄素+DDP-40(20μmol/L姜黄素+40μmol/L顺铂)、姜黄素+DDP-50(20μmol/L姜黄素+50μmol/L顺铂)、姜黄素+DDP-60组(20μmol/L姜黄素+60μmol/L顺铂),CCK-8法检测细胞活性,计算顺铂半数抑制浓度(IC 50),筛选顺铂和姜黄素的合适作用浓度。(2)将OVCAR-3细胞分为对照组(常规处理)、DDP组(20.5μg/mL顺铂)、姜黄素组(20μmol/L姜黄素)、姜黄素+DDP组(20μmol/L姜黄素+20.5μg/mL顺铂);将OVCAR-3/DDP细胞分为对照组(常规处理)、DDP组(42.1μg/mL顺铂)、姜黄素组(20μmol/L姜黄素)、姜黄素+DDP组(20μmol/L姜黄素+42.1μg/mL顺铂),采用流式细胞术检测细胞凋亡率,实时荧光定量(qRT)-PCR检测miR-135b-5p表达,qRT-PCR和免疫印迹法分别检测瓣状核酸内切酶-1(flap endonuclease 1,FEN1)mRNA和蛋白表达。结果:相同顺铂浓度时,与DDP组比较,姜黄素+DDP联合组OVCAR-3和OVCAR-3/DDP细胞活性明显降低(P均<0.001)。DDP作用于OVCAR-3和
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(1μmol/L cisplatin),DDP-5(5μmol/L cisplatin),DDP-10(10μmol/L cisplatin),DDP-15(15μmol/L cisplatin),DDP-20(20μmol/L cisplatin),DDP-30(30μmol/L cisplatin),curcumin+DDP-1(20μmol/L curcumin+1μmol/L cisplatin),curcumin+DDP-5(20μmol/L curcumin+5μmol/L cisplatin),curcumin+DDP-10(20μmol/L curcumin+10μmol/L cisplatin),curcumin+DDP-15(20μmol/L curcumin+15μmol/L cisplatin),curcumin+DDP-20(20μmol/L curcumin+20μmol/L cisplatin)and curcumin+DDP-30 groups(20μmol/L curcumin+30μmol/L cisplatin).OVCAR-3/DDP cells were divided into Control,DDP-10(10μmol/L cisplatin),DDP-20(20μmol/L cisplatin),DDP-30(30μmol/L cisplatin),DDP-40(40μmol/L cisplatin),DDP-50(50μmol/L cisplatin),DDP-60(60μmol/L cisplatin),curcumin+DDP-10(20μmol/L curcumin+10μmol/L cisplatin),curcumin+DDP-20(20μmol/L curcumin+20μmol/L cisplatin),curcumin+DDP-30(20μmol/L curcumin+30μmol/L cisplatin),curcumin+DDP-40(20μmol/L curcumin+40μmol/L cisplatin),curcumin+DDP-50(20μmol/L curcumin+50μmol/L cisplatin),curcumin+DDP-60(20μmol/L curcumin+60μmol/L cisplatin)groups.CCK-8 method was used to detect the cell viability,the median inhibitory concentration of cisplatin(IC 50)was calculated,and the appropriate concentration of cisplatin and curcumin was screened.(2)OVCAR-3 cells were divided into Control group,DDP group(20.5μg/mL cisplatin),curcumin group(20μmol/L curcumin)and curcumin+DDP group(20μmol/L curcumin+20.5μg/mL cisplatin).OVCAR-3/DDP cells were divided into Control group,DDP group(42.1μg/mL cisplatin),curcumin group(20μmol/L curcumin)and curcumin+DDP group(20μmol/L curcumin+42.1μg/mL cisplatin).The apoptosis rate was detected by flow cytometry,the expression of miR-135b-5p was detec
作者
马蓉
李娟
王芳
MA Rong;LI Juan;WANG Fang(Department of Gynaecology,Xinjiang Uygur Autonomous Region Hospital of Traditional Chinese Medicine,Urumqi Xinjiang 830099,China)
出处
《江苏大学学报（医学版）》
CAS
2024年第4期324-330,共7页
Journal of Jiangsu University：Medicine Edition
基金
新疆维吾尔自治区自然科学基金资助项目(2021D01C237)。
