Xu Yongqian, Zheng Yujie, Duan Yan, Ma Lin, Nan Ping
Department of Obstetrics and Gynecology, Shengli Oilfield Central Hospital, Dongying, Shandong 257000, P.R. China.
Oncol Lett. 2021 May;21(5):392. doi: 10.3892/ol.2021.12653. Epub 2021 Mar 18.
Cervical cancer (CC), also known as invasive cervical carcinoma, is one of the most common gynecologic malignancies. The aim of the present study was to investigate the function of microRNA (miR)-125a-5p on CC progression and cisplatin (DDP) resistance. For this purpose, reverse transcription-quantitative PCR (RT-qPCR) was used to assess the expression of miR-125a-5p and LIMK1 in CC tissues, corresponding normal tissues and cells (human CC cell lines: C-33A, CaSKi; human cervical epithelial cells: HUCEC). Cisplatin (DDP) resistant cervical cancer cell lines were established (C-33A/DDP and CaSKi/DDP cell lines). RT-qPCR results demonstrated that miR-125a-5p or LIM kinase 1 (LIMK1) expression was downregulated or upregulated in C-33A/DDP and CaSKi/DDP cells, respectively. MTT assay, flow cytometry analysis and Western blotting were employed to detect the proliferation, apoptosis rate, IC of DDP and the expression of drug resistance-related proteins (P-glycoprotein and glutathione S-transferase-π). The targeting relationship between miR-125a-5p and LIMK1 was confirmed by the TargetScan database and dual-luciferase reporter gene assay. In CC tissues and cell lines, compared with normal tissues or HUCEC, miR-125a-5p expression was downregulated and LIMK1 expression was upregulated. The transfection with miR-125a-5p mimics decreased the proliferation of CaSKi/DDP cells, increased the apoptosis rate, reduced the IC of DDP, and downregulated the expression of drug resistance-related proteins; conversely, LIMK1 overexpression decreased the apoptosis rate, increased the IC of DDP, and upregulated the expression of drug resistance-related proteins. The luciferase reporter gene assay demonstrated that miR-125a-5p targeted and negatively regulated LIMK1. miR-125a-5p could partially reverse the effect of LIMK1 on the proliferation, apoptosis, IC of DDP and the expressions of drug resistance-related proteins. The findings of the present study indicated that miR-125a-5p sensitizes CC cells to DDP by targeting LIMK1, hence increasing the anticancer efficacy of cisplatin.
宫颈癌(CC),也称为浸润性宫颈癌,是最常见的妇科恶性肿瘤之一。本研究的目的是探讨微小RNA(miR)-125a-5p在宫颈癌进展和顺铂(DDP)耐药中的作用。为此,采用逆转录定量聚合酶链反应(RT-qPCR)检测miR-125a-5p和LIMK1在宫颈癌组织、相应正常组织及细胞(人宫颈癌细胞系:C-33A、CaSKi;人宫颈上皮细胞:HUCEC)中的表达。建立顺铂(DDP)耐药宫颈癌细胞系(C-33A/DDP和CaSKi/DDP细胞系)。RT-qPCR结果显示,在C-33A/DDP和CaSKi/DDP细胞中,miR-125a-5p表达下调,而LIM激酶1(LIMK1)表达上调。采用MTT法、流式细胞术分析和蛋白质印迹法检测细胞增殖、凋亡率、DDP的半数抑制浓度(IC)以及耐药相关蛋白(P-糖蛋白和谷胱甘肽S-转移酶-π)的表达。通过TargetScan数据库和双荧光素酶报告基因实验证实了miR-125a-5p与LIMK1之间的靶向关系。在宫颈癌组织和细胞系中,与正常组织或HUCEC相比,miR-125a-5p表达下调,LIMK1表达上调。转染miR-125a-5p模拟物可降低CaSKi/DDP细胞的增殖,增加凋亡率,降低DDP的IC,并下调耐药相关蛋白的表达;相反,过表达LIMK1可降低凋亡率,增加DDP的IC,并上调耐药相关蛋白的表达。荧光素酶报告基因实验表明,miR-125a-5p靶向并负调控LIMK1。miR-125a-5p可部分逆转LIMK1对细胞增殖、凋亡、DDP的IC及耐药相关蛋白表达的影响。本研究结果表明,miR-125a-5p通过靶向LIMK1使宫颈癌细胞对DDP敏感,从而提高顺铂的抗癌疗效。