MicroRNA (miRNA) 是单个 RNA 分子，在哺乳动物细胞中充当基因表达的全局调节因子，因此成为治疗癌症的有吸引力的靶标。在这里，我们的目的是研究 miRNA-141 (miR-141) 在宫颈癌中的可能参与，并确定其在宫颈癌细胞系中的潜在靶标。HeLa 和 C-33A 细胞中 miR-141 的水平已使用实时定量 PCR (qRT-PCR)。 新的 miR-141 构建体已在带有 GFP 标记的 CMV 启动子载体中进行。通过微阵列分析，我们鉴定了转染的 HeLa 细胞中 miR-141 潜在调控的基因。在用抑制剂、拮抗剂 miR-141 或 miR-141 过表达转染的 HeLa 细胞中研究了杀伤样受体 C1 (KLRC1)、KLRC3、癌胚抗原相关细胞粘附分子 3 (CAM3) 和 CAM6 的蛋白质谱使用免疫印迹和流式细胞术检测载体。最后，使用ELISA测定法监测转染的HeLa细胞产生的细胞因子。与正常宫颈HCK1T细胞系相比，HeLa和C-33A细胞中miR-141的表达显着增加。与 miR-141 过表达载体的转染不同，用抑制剂拮抗剂 miR-141 转染 HeLa 细胞对癌细胞活力显示出有效的影响。过表达 miR-141 的 HeLa 细胞的微阵列数据提供了一百个下调基因，包括 KLRC1、KLRC3、CAM3 和 CAM6。在转染 miR-141 过表达的 HeLa 细胞中，KLRC1 和 KLRC3 表达谱显着减少，同时白细胞介素 8 (IL-8) 减少，表明 miR-141 在避免 HeLa 细胞中程序性细胞死亡中的作用。同样，在 miR-141 转导的细胞中，CAM3 和 CAM6 表达显着降低，伴随着转化生长因子 β (TGF-β) 水平的增加，表明 miR-141 对癌细胞迁移的影响。 IntaRNA 程序和 miRWalk 用于检查 miR-141 和已识别基因之间的直接相互作用和潜在结合位点。基于此，每个潜在靶标的播种区域被克隆到 pGL3 对照载体中荧光素酶报告基因的上游。有趣的是，构建的载体的荧光素酶活性在预转染 miR-141 过表达载体的 HeLa 细胞中显着降低，而在预转染 miR-141 特异性抑制剂的细胞中大幅增加。 总之，这些数据揭示了一种有效的 miR-141-基于支持宫颈癌进展的机制，并将 miR-141 确定为可靠的治疗靶点。© 2024。作者。

MicroRNAs (miRNAs) are single RNA molecules that act as global regulators of gene expression in mammalian cells and thus constitute attractive targets in treating cancer. Here we aimed to investigate the possible involvement of miRNA-141 (miR-141) in cervical cancer and to identify its potential targets in cervical cancer cell lines.The level of miR-141 in HeLa and C-33A cells has been assessed using the quantitative real-time PCR (qRT-PCR). A new miR-141 construct has been performed in a CMV promoter vector tagged with GFP. Using microarray analysis, we identified the potentially regulated genes by miR-141 in transfected HeLa cells. The protein profile of killer-like receptor C1 (KLRC1), KLRC3, carcinoembryonic antigen-related cell adhesion molecule 3 (CAM3), and CAM6 was investigated in HeLa cells transfected with either an inhibitor, antagonist miR-141, or miR-141 overexpression vector using immunoblotting and flow cytometry assay. Finally, ELISA assay has been used to monitor the produced cytokines from transfected HeLa cells.The expression of miR-141 significantly increased in HeLa and C-33A cells compared to the normal cervical HCK1T cell line. Transfection of HeLa cells with an inhibitor, antagonist miR-141, showed a potent effect on cancer cell viability, unlike the transfection of miR-141 overexpression vector. The microarray data of HeLa cells overexpressed miR-141 provided a hundred of downregulated genes, including KLRC1, KLRC3, CAM3, and CAM6. KLRC1 and KLRC3 expression profiles markedly depleted in HeLa cells transfected with miR-141 overexpression accompanied by decreasing interleukin 8 (IL-8), indicating the role of miR-141 in avoiding programmed cells death in HeLa cells. Likewise, CAM3 and CAM6 expression reduced markedly in miR-141 transduced cells accompanied by an increasing level of transforming growth factor beta (TGF-β), indicating the impact of miR-141 in cancer cell migration. The IntaRNA program and miRWalk were used to check the direct interaction and potential binding sites between miR-141 and identified genes. Based on this, the seeding regions of each potential target was cloned upstream of the luciferase reporter gene in the pGL3 control vector. Interestingly, the luciferase activities of constructed vectors were significantly decreased in HeLa cells pre-transfected with miR-141 overexpression vector, while increasing enormously in cells pre-transfected with miR-141 specific inhibitor.Together, these data uncover an efficient miR-141-based mechanism that supports cervical cancer progression and identifies miR-141 as a credible therapeutic target.© 2024. The Author(s).