miR-193b-5p靶向CD44v6调控宫颈癌细胞侵袭、迁移以及上皮-间质转化的机制研究  

Study on the mechanism of miR⁃193b⁃5p targeting CD44v6 to regulate the invasion,migration and epithelial⁃mesenchymal transition of cervical cancer cells

作　　者：热孜宛古丽·约麦尔 买买提·司马义 张晓玲[1] 叶勒丹·马汉 周玲[1] REZIWANGULI Yuemaier;MAIMAITI Simayi;ZHANG Xiaoling;YELEDAN Mahan;ZHOU Ling(Medical Research and Transformation Center,People′s Hospital of Xinjiang Uygur Autonomous Region,Urumqi 830001,China;Pharmacy Department of the First Affiliated Hospital of Xinjiang Medical University,Urumqi 830001,China)

机构地区：[1]新疆维吾尔自治区人民医院科研教育中心-医学研究与转化管理科,新疆乌鲁木齐830001 [2]新疆医科大学第一附属医院药学部,新疆乌鲁木齐830011

出　　处：《海南医科大学学报》2025年第7期522-530,共9页Journal of Hainan Medical University

基　　金：新疆维吾尔自治区自然科学基金(2022D01C104)。

摘　　要：目的:探究微小RNA-193b-5p(miR-193b-5p)对宫颈癌的影响及作用机制。方法:收集38例宫颈癌患者手术切除的宫颈癌肿瘤组织及同期38例来源于我院因良性病变切除全子宫患者的正常宫颈组织,RT-qPCR检测两种组织中miR-193b-5p表达水平;将SiHa细胞分为对照组、mimics NC组、miR-193b-5p mimics组、inhibitorsNC组、miR-193b-5pinhibitors组,进行相应转染后,RT-qPCR检测各组细胞中miR-193b-5p表达水平,CCK-8法、细胞划痕实验、Transwell小室法分别检测各组SiHa细胞的增殖活性、迁移、侵袭,细胞免疫荧光染色和Western Blot检测各组SiHa细胞中E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)等上皮-间质转化(EMT)相关蛋白表达,生物信息学结合双荧光素酶报告基因实验预测和验证miR-193b-5p与白细胞分化抗原44号拼接变异体v6(CD44v6)的靶向调控关系。结果:宫颈癌组织中miR-193b-5p相对表达量显著低于与正常宫颈组织(P<0.05)。与对照组、mimics NC组比较,miR-193b-5p mimics组SiHa细胞中miR-193b-5p相对表达量显著上调(P<0.05),细胞增殖活性、细胞划痕愈合率显著降低(P<0.05),细胞侵袭数目显著减少(P<0.05),E-cadherin荧光染色强度和蛋白相对表达量显著增加(P<0.05),N-cadherin、Vimentin荧光染色强度和蛋白相对表达量显著减少(P<0.05);而与对照组、inhibitors NC组比较,miR-193b-5p inhibitors组SiHa细胞中miR-193b-5p相对表达量显著下调(P<0.05),细胞增殖活性、细胞划痕愈合率显著升高(P<0.05),细胞侵袭数目显著增加(P<0.05),E-cadherin荧光染色强度和蛋白相对表达量显著减少(P<0.05),N-cadherin、Vimentin荧光染色强度和蛋白相对表达量显著增加(P<0.05)。miR-193b-5p与CD44v6的3′-非翻译区(3′-UTR)存在结合位点,且与mimics NC组比较,将miR-193b-5p mimics转染至SiHa细胞中显著抑制了CD44v6野生型(WT)的相对荧光素酶活性(P<0.05)。结论:miR-193b-5p在人宫颈癌肿瘤组织�Objective:To explore the regulatory effect of microRNA-193b-5p(miR-193b-5p)on the biological behavior of human cervical cancer SiHa cells and its molecular mechanism.Methods:Cervical cancer tissues from 38 patients with cervical cancer and normal cervical tissues from 38 patients with hysterectomy during the same period were collected,RT-qPCR was used to detect the expression levels of miR-193b-5p in the two tissues.SiHa cells were divided into the control group,the mimics NC group,the miR-193b-5p mimics group,the inhibitors NC group and the miR-193b-5p inhibitors group,after corresponding transfection,the expression levels of miR-193b-5p in the cells of each group were detected by RT-qPCR,the proliferation activity,migration and invasion of SiHa cells in each group were detected by CCK-8 method,cell scratch assay and Transwell chamber method,respectively,cell immunofluorescence staining and Western Blot was used to detect the expression of E-cadherin,N-cadherin,Vimentin and other epithelial-mesenchymal transition(EMT)-related proteins in SiHa cells of each group,bioinformatics combined with dual luciferase reporter gene experiments predicted and verified the targeted regulatory relationship between miR-193b-5p and leukocyte differentiation antigen 44 splicing variant v6(CD44v6).Results:The relative expression of miR-193b-5p in cervical cancer tissues was significantly lower than that in normal cervical tissues(P<0.05).Compared with the control group and the mimics NC group,the relative expression of miR-193b-5p in SiHa cells in the miR-193b-5p mimics group was significantly upregulated(P<0.05),the cell proliferation activity and cell scratch healing rate were significantly reduced(P<0.05),the number of cell invasion was significantly reduced(P<0.05),the fluorescence staining intensity and relative protein expression of E-cadherin were significantly increased(P<0.05),and the fluorescence staining intensity and relative protein expression of N-cadherin and Vimentin were significantly decreased(P<0.05);compared wit

关 键 词：宫颈癌 miR-193b-5p CD44V6 侵袭 迁移 上皮-间质转化 

分 类 号：R737.33[医药卫生—肿瘤]