miR-195-5p通过靶向STAT3调控免疫抑制基因PD-L1参与卵巢癌增殖和侵袭的机制研究  

miR-195-5p suppresses the proliferation and invasion of ovarian cancer by targeting STAT3 and regulating the immunosuppressive gene PD-L1

作　　者：陈丽[1] 张静怡 王露玉 孙白云 郭亮生[1] CHEN Li;ZHANG Jingyi;WANG Luyu;SUN Baiyun;GUO Liangsheng(Department of Obstetrics and Gynecology,Second Affiliated Hospital of Soochow University,Suzhou 215000,China)

机构地区：[1]苏州大学附属第二医院妇产科,215000

出　　处：《免疫学杂志》2020年第7期572-577,共6页Immunological Journal

摘　　要：目的探究微小RNA(microRNA,miR)-195-5p通过靶向STAT3调控免疫抑制基因程序性死亡受体-配体1(programmed cell death-ligand 1,PD-L1)参与卵巢癌增殖和侵袭的机制。方法通过双荧光素酶报告基因检测方法验证miR-195-5p靶向STAT3。将卵巢癌细胞系SKOV-3分为4组:对照组、mimic组、mimic+STAT3组和STAT3组。通过质粒转染技术过表达miR-195-5p和STAT3。qPCR和Western blot检测RNA或蛋白的表达水平。分别通过CCK-8法、Transwell实验检测各组的细胞活力、侵袭能力。通过荷瘤裸鼠实验在体内验证过表达miR-195-5p对肿瘤生长、STAT3和PD-L1的影响。结果 miR-195-5p直接靶向STAT3。过表达miR-195-5p抑制STAT3 mRNA和蛋白的表达水平(P<0.05),过表达STAT3显著逆转miR-195-5p对STAT3的抑制作用(P<0.05)。Mimic组的细胞活力和细胞迁移能力显著低于对照组(P<0.05),STAT3组的细胞活力和细胞迁移能力显著高于对照组(P<0.05),并且mimic+STAT3组的细胞活力和细胞迁移能力显著高于mimic组(P<0.05)。荷瘤裸鼠实验显示miR-195-5p组荷瘤裸鼠的肿瘤体积和质量均显著低于对照组(P<0.05)。miR-195-5p组荷瘤裸鼠的肿瘤组织中STAT3和PD-L1蛋白水平显著低于对照组(P<0.05)。结论 miR-195-5p可通过靶向STAT3抑制PD-L1的表达并抑制卵巢癌细胞生长、侵袭和免疫抑制反应。This study was designed to explore the mechanism of microRNA(miR)-195-5 p involved in the proliferation and invasion of ovarian cancer by targeting STAT3 to regulate the immunosuppressive gene programmed death cell-ligand 1(PD-L1).Dual luciferase reporter assay was used to verify that miR-195-5 p targets STAT3.The cancer cell line SKOV-3 was divided into four groups:control group,mimic group,mimic+STAT3 group and STAT3 group.miR-195-5 p and STAT3 were overexpressed by plasmid transfection techniques.qPCR and Western blot were used respectively to detect the expression level of RNA and protein.The cell viability and invasive ability of each group were detected by CCK-8 method and Transwell test,respectively;the effect of miR-195-5 p overexpression on tumor growth,STAT3 and PD-L1 were verified in vivo by using tumor-bearing nude mice.DatashowedthatmiR-195-5 pdirectlytargetsSTAT3.OverexpressionofmiR-195-5 pinhibitedtheexpression levels of STAT3 mRNA and protein,while overexpression of STAT3 significantly reversed the inhibitory effect of miR-195-5 p on STAT3.The cell viability and cell migration ability of the mimic group were significantly lower than those of the control group(P<0.05).The cell viability and cell migration ability of STAT3 group were significantly higher than that of the control group(P<0.05).The cell viability and cell migration ability of the mimic+STAT3 group were significantly higher than those of the mimic group(P<0.05).Tumor-bearing nude mice showed that the tumor volume and quality of the tumor-bearing nude mice in the miR-195-5 p group were significantly lower than those in the control group(P<0.05).The levels of STAT3 and PD-L1 proteins in tumor tissues of miR-195-5 p tumor-bearing nude mice were significantly lower than those in the control group(P<0.05).Taken together,miR-195-5 p can inhibit the expression of PD-L1 and suppress the growth,invasion and immunosuppressive response of ovarian cancer cells by targeting STAT3.

关 键 词：卵巢癌 微小RNA miR-195-5p 免疫抑制 程序性死亡受体-配体1 

分 类 号：R73[医药卫生—肿瘤]