Zhang Wenfang, Han Dongyan
Department of Obstetrics and Gynecology, Huangshi Central Hospital, E'dong Healthcare Group, Huangshi, 435000 Hubei China.
Department of Obstetrics and Gynecology, Kaifeng Third People's Hospital, Kaifeng, 475000 Henan China.
Cytotechnology. 2021 Aug;73(4):585-592. doi: 10.1007/s10616-021-00479-y. Epub 2021 Jun 8.
Numerous studies have found that microRNAs (miRNAs) are involved in regulating various tumor-related biological functions. The downregulation of miR185-3p have been identified in various types of cancer but the effect and its underlying molecular mechanism in cervical cancer have not been elucidated. Therefore, it is important to investigate the role of miRNAs associated with cervical cancer and its corresponding molecular mechanism to develop new therapeutic targets. The cell counting kit (CCK-8) assay was performed to measure the cell viability. The quantitative real-time PCR (qRT-PCR) and western blot analyses were carried out to identify mRNA and protein expression levels, respectively. Besides, a luciferase activity assay was conducted to confirm the target miRNA gene predictions. In this study, it is found that miR185-3p expression was potentially downregulated in cervical cancer tissues when compared with normal tissues. The CCK-8 results indicated that miR185-3p overexpression suppressed the cancer cell proliferation and the downregulation of miR185-3p enhanced the cancer cell growth. Further, enhanced miR185-3p expression led to a reduction in Annexin-A8 (Anx-A8) expression but miR185-3p inhibition promoted ANX-A8 levels in cervical cancer cells. The luciferase reporter assay indicated that ANX-A8 was a direct target of miR185-3p in cervical cancer cells.
众多研究发现,微小RNA(miRNA)参与调控多种肿瘤相关生物学功能。在各类癌症中均已发现miR185 - 3p表达下调,但它在宫颈癌中的作用及其潜在分子机制尚未阐明。因此,研究与宫颈癌相关的miRNA的作用及其相应分子机制对于开发新的治疗靶点具有重要意义。采用细胞计数试剂盒(CCK - 8)法检测细胞活力。分别通过定量实时聚合酶链反应(qRT - PCR)和蛋白质免疫印迹分析来鉴定mRNA和蛋白质表达水平。此外,进行荧光素酶活性测定以确认miRNA靶基因预测结果。本研究发现,与正常组织相比,宫颈癌组织中miR185 - 3p的表达可能下调。CCK - 8结果表明,miR185 - 3p过表达抑制癌细胞增殖,而miR185 - 3p下调则促进癌细胞生长。此外,miR185 - 3p表达增强导致宫颈癌细胞中膜联蛋白A8(Anx - A8)表达降低,但抑制miR185 - 3p则促进宫颈癌细胞中ANX - A8水平升高。荧光素酶报告基因测定表明,ANX - A8是宫颈癌细胞中miR185 - 3p的直接靶标。