miR-381-3p调控PD-L1对卵巢癌细胞卡铂化疗耐药的影响

摘要/Abstract

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通过调控PD-L1参与对卵巢癌细胞的卡铂耐药，为卵巢癌耐药治疗提供参考依据。

关键词: 微小RNA , -381-3p；程序性死亡配体-1；卵巢癌细胞；卡铂

Abstract:

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 intervention 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 miR-381-3p analog PD-L1 empty vector intervention group， while the proportion of S-stage cells was lower than that of the co-transfected miR-381-3p analog PD-L1 empty vector intervention group （P< 0.05）.The apoptosis rate of the miR-381-3p analog intervention group was higher than that of the miR-381-3p negative intervention group， and the apoptosis rate of the co-transfected miR-381-3p analog and the PD-L1 overexpressed plasmid intervention group was lower than that of the co-transfected miR-381-3p analog PD-L1 empty vector intervention group （P<0.05）. The expression of PD-L1， MDR1， and Cyclin D1 proteins in the miR-381-3p analog intervention group was lower than that of the miR-381-3p negative intervention group， while the expression of the cleaved-caspase-3 protein was higher than that of the miR-381-3p negative intervention group （P<0.05）. The expression of cleaved-caspase-3 protein in the co-transfected miR-381-3p analog and PD-L1 overexpressed plasmid intervention group was lower than that of the co-transfected miR-381-3p analog PD-L1 empty vector intervention group （P<0.05）.Conclusion： miR-381-3p regulates PD-L1， which is involved in carboplatin resistance of ovarian cancer cells. It provides a reference for the treatment of drug resistance in ovarian cancer.

Key words: miR-381-3p； , Programmed death ligand-1； , Ovarian cancer cells； , Carboplatin

中图分类号:

R737.31

徐洁欢, 邓玉屏, 李耀军, 廉萍, 叶枫.

miR-381-3p调控PD-L1对卵巢癌细胞卡铂化疗耐药的影响 [J]. 中国医药导刊, 2024, 26(9): 929-936.

XU Jiehuan, DENG Yuping, LI Yaojun, LIAN Ping, YE Feng.

Effects of miR-381-3p Regulating PD-L1 on the Resistance of Carboplatin Chemotherapy in Ovarian Cancer Cells [J]. CHINESE JOURNAL OF MEDICINAL GUIDE, 2024, 26(9): 929-936.

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