Department of Obstetrics and Gynecology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu 210036, P.R. China.
Department of Radiology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu 210036, P.R. China.
Oncol Rep. 2018 Jun;39(6):3015-3024. doi: 10.3892/or.2018.6391. Epub 2018 Apr 23.
Cervical cancer is the third most common type of cancer in women, and microRNAs play an important role in this type of cancer. The elevated expression of miR-146a is involved in the pathogenesis of cancers generally, but its role in cervical cancer has not been fully elucidated. In the present study, we assessed the expression of miR-146a in G>C polymorphisms and confirmed that the overexpression of miR-146a promoted cervical cancer cell viability. The recombinant expression plasmids pre-miR-146a-G or pre-miR-146a-C including single nucleotide polymorphisms (SNP) were successfully constructed. Pre-miR-146a-G or pre-miR-146a-C was transfected into cervical cancer cells or immortalized non-tumorigenic cells and the expression of miR-146a was evaluated by real-time PCR. The cell viability, cell-cycle analysis and apoptosis were assessed using Cell Counting Kit-8 assay (CCK-8), flow cytometry and cleaved caspase-3 protein expression, respectively. The expression of interleukin 1 receptor associated kinase 1 (IRAK1), TNF receptor-associated factor 6 (TRAF6) and cyclin D1 was assessed following the transfection with a miR-146a mimic or a negative control. The cell viability and the number of S-phase cells increased after transfection with miR-146a mimic or an IRAK1 or TRAF6 interference fragment. After transfection, IRAK1 and TRAF6 protein expression was downregulated and the expression of cyclin D1 was upregulated, however apoptosis and cleaved caspase-3 were not affected. Polymorphisms in miR-146a precursor may be linked to the expression of miR-146a and may be a potential target for cervical cancer therapy.
宫颈癌是女性中第三常见的癌症类型，而 microRNAs 在这种癌症中发挥着重要作用。miR-146a 的表达升高通常与癌症的发病机制有关，但它在宫颈癌中的作用尚未完全阐明。在本研究中，我们评估了 miR-146a 在 G>C 多态性中的表达，并证实 miR-146a 的过表达促进了宫颈癌细胞的活力。成功构建了包含单核苷酸多态性（SNP）的重组表达质粒 pre-miR-146a-G 或 pre-miR-146a-C。将 pre-miR-146a-G 或 pre-miR-146a-C 转染到宫颈癌细胞或永生化非肿瘤细胞中，并通过实时 PCR 评估 miR-146a 的表达。使用 Cell Counting Kit-8 assay (CCK-8)、流式细胞术和 cleaved caspase-3 蛋白表达分别评估细胞活力、细胞周期分析和细胞凋亡。转染 miR-146a 模拟物或阴性对照后，评估白细胞介素 1 受体相关激酶 1 (IRAK1)、肿瘤坏死因子受体相关因子 6 (TRAF6) 和细胞周期蛋白 D1 的表达。转染 miR-146a 模拟物或 IRAK1 或 TRAF6 干扰片段后，细胞活力和 S 期细胞数量增加。转染后，IRAK1 和 TRAF6 蛋白表达下调，cyclin D1 表达上调，但细胞凋亡和 cleaved caspase-3 不受影响。miR-146a 前体的多态性可能与 miR-146a 的表达有关，可能成为宫颈癌治疗的潜在靶点。
