摘要
目的:探讨miR-381-3p调控PD-L1对卵巢癌细胞卡铂化疗耐药的影响。方法:采用实时荧光定量PCR检测比较SKOV3细胞及SKOV3/CBP细胞中PD-L1、miR-381-3p的表达情况。细胞转染miR-381-3p模拟物和抑制物,分组如下:SKOV3/CBP miR-381-3p阴性对照组,SKOV3/CBP miR-381-3p模拟物组,SKOV3 miR-381-3p阴性对照组,SKOV3 miR-381-3p抑制物组,实时荧光定量PCR检测卵巢癌卡铂耐药细胞中miR-381-3p对PD-L1表达的影响。细胞转染miR-381-3p模拟物和PD-L1过表达质粒卡铂干预处理,分组如下:miR-381-3p阴性干预组,miR-381-3p模拟物干预组,共转染miR-381-3p模拟物PD-L1空载体干预组,共转染miR-381-3p模拟物与PD-L1过表达质粒干预组,流式细胞术检测miR-381-3p调控PD-L1表达对细胞周期和凋亡变化,蛋白质印迹法检测miR-381-3p调控PD-L1表达对MDR1、Cyclin D1和cleaved-caspase-3的表达变化。结果:卵巢癌卡铂耐药SKOV3细胞组中PD-L1表达水平高于卵巢癌SKOV3组,而miR-381-3p表达水平低于卵巢癌SKOV3组(P<0.05);SKOV3 miR-381-3p抑制物组中PD-L1的表达高于SKOV3 miR-381-3p阴性对照组,SKOV3/CBP miR-381-3p模拟物组中PD-L1的表达低于SKOV3/CBP miR-381-3p阴性对照组(P<0.05);共转染miR-381-3p模拟物与PD-L1过表达质粒干预组的G1期细胞比例高于共转染miR-381-3p模拟物PD-L1空载体干预组,而S期细胞比例低于共转染miR-381-3p模拟物PD-L1空载体干预组(P<0.05)。miR-381-3p模拟物干预组细胞凋亡率高于miR-381-3p阴性干预组,共转染miR-381-3p模拟物与PD-L1过表达质粒干预组细胞凋亡率低于共转染miR-381-3p模拟物PD-L1空载体干预组(P<0.05)。miR-381-3p模拟物干预组中PD-L1、MDR1、Cyclin D1蛋白表达低于miR-381-3p阴性干预组,而其cleaved-caspase-3蛋白表达高于miR-381-3p阴性干预组(P<0.05)。共转染miR-381-3p模拟物与PD-L1过表达质粒干预组中cleaved-caspase-3蛋白表达低于共转染miR-381-3p模拟物PD-L1空载体干预组(P<0.05)。结论:miR-381-3p通过调�
Objective:To explore the effect of miR-381-3p on the regulation of PD-L1 on the resistance of carboplatin chemotherapy in ovarian cancer cells.Methods:Real-time fluorescence quantitative PCR was used to detect and compare the expression of PD-L1 and miR-381-3p in SKOV3 cells and SKOV3/CBP cells.Cell transfection mimetics and inhibitors of miR-381-3p are grouped as follows:SKOV3/CBP miR-381-3p negative control group,SKOV3/CBP miR-381-3p analog group,SKOV3 miR-381-3p negative control group,SKOV3 miR-381-3p inhibitor group.Real-time fluorescence quantitative PCR was used to detece the effect of miR-381-3p on PD-L1 expression in carboplatin-resistant cells of ovarian cancer.Cells were transfected with miR-381-3p analog and PD-L1 overexpression plasmid for carboplatin intervention treatment,grouped as follows:miR-381-3p negative intervention group,miR-381-3p analog inter-vention group,co-transfection of miR-381-3p analog PD-L1 empty vector intervention group,co-transfection of miR-381-3p analog and PD-L1 overexpression plasmid intervention group.Flow cytometry was used to detect the effects in the cell cycle and apoptosis regulated by miR-381-3p on PD-L1 expression,and Western blotting was used to detect the expression changes of MDR1,Cyclin D1 and cleaved-caspase-3 regulated by miR-381-3p on PD-L1 expression.Results:The expression level of PD-L1 in the carboplatin-resistant SKOV3 cell group of ovarian cancer was higher than that of the SKOV3 cell group of ovarian cancer,while the expression level of miR-381-3p was lower than that of the SKOV3 cell group of ovarian cancer(P<0.05).The expression of PD-L1 in the SKOV3 miR-381-3p inhibitor group was higher than that of the SKOV3 miR-381-3p negative control group,and the expression of PD-L1 in the SKOV3/CBP miR-381-3p analog group was lower than that of the SKOV3/CBP miR-381-3p negative control group(P<0.05).The proportion of G1-stage cells in the co-transfected miR-381-3p analog and PD-L1 overexpressed plasmid intervention group was higher than that of the co-transfected
作者
徐洁欢
邓玉屏
李耀军
廉萍
叶枫
XU Jiehuan;DENG Yuping;LI Yaojun;LIAN Ping;YE Feng(Department of Gynecology and Pediatrics,Changsha Health Vocational College,Hunan Changsha 410600,China;The Fifth Department of Gynecological Tumor,the Affiliated Cancer Hospital of Xiangya School of Medicine,Hunan Changsha 410013,China)
出处
《中国医药导刊》
2024年第9期929-936,共8页
Chinese Journal of Medicinal Guide
基金
湖南省自然科学基金(2022JJ60110)。
