Oboshi Wataru, Hayashi Keisuke, Takeuchi Hiroaki, Ikeda Katsuhide, Yamaguchi Yoshitaka, Kimura Asako, Nakamura Takehiro, Yukimasa Nobuyasu
Department of Medical Technology and Sciences, International University of Health and Welfare, Narita Chiba 286-8686, Japan.
Department of Clinical Laboratory, Shikoku Central Hospital, Shikokuchuo, Ehime 799-0193, Japan.
Oncol Lett. 2020 Nov;20(5):210. doi: 10.3892/ol.2020.12073. Epub 2020 Sep 8.
MicroRNAs (miRNAs) exert critical roles in the majority of biological and pathological processes. Recent studies have associated miR-150 with a number of different cancer types. However, little is known about miR-150 targets in cervical cancer. In the present study, the HeLa human cervical cancer cell line was transfected with hsa-miR-150-5p mimics, hsa-miR-150-5p inhibitors or miRNA controls. miR-150 was predicted to bind the 3'untranslated region (3'UTR) of the gene, which encodes the cyclin-dependent kinase inhibitor 1B (p27). The direct binding between miR-150 and the 3'UTR of was confirmed using dual-luciferase reporter assays. The effects of miR-150 on mRNA expression, p27 protein expression, cell cycle and cell proliferation were determined using reverse-transcription quantitative PCR, western blot analysis, flow cytometry and WST-8 assays, respectively. miR-150 was demonstrated to directly target the 3'UTR of in transfected HeLa cells. The expression of mRNA and p27 protein was reduced by miR-150 mimics, and increased by miR-150 inhibitors. Moreover, the overexpression of miR-150 promoted cell cycle progression from the G0/G1 to the S phase and led to a significant increase in HeLa cell proliferation. The results of the present study indicated that miR-150 promotes HeLa cell cycle progression and proliferation via the suppression of p27 expression.
微小RNA(miRNA)在大多数生物和病理过程中发挥着关键作用。最近的研究将miR-150与多种不同类型的癌症联系起来。然而,关于miR-150在宫颈癌中的靶标知之甚少。在本研究中,将hsa-miR-150-5p模拟物、hsa-miR-150-5p抑制剂或miRNA对照转染到人宫颈癌HeLa细胞系中。预测miR-150与编码细胞周期蛋白依赖性激酶抑制剂1B(p27)的基因的3'非翻译区(3'UTR)结合。使用双荧光素酶报告基因测定法证实了miR-150与该基因3'UTR之间的直接结合。分别使用逆转录定量PCR、蛋白质印迹分析、流式细胞术和WST-8测定法确定了miR-150对该基因mRNA表达、p27蛋白表达、细胞周期和细胞增殖的影响。结果表明,在转染的HeLa细胞中,miR-150直接靶向该基因的3'UTR。miR-150模拟物降低了该基因mRNA和p27蛋白的表达,而miR-150抑制剂则使其增加。此外,miR-150的过表达促进了细胞周期从G0/G1期向S期的进展,并导致HeLa细胞增殖显著增加。本研究结果表明,miR-150通过抑制p27表达促进HeLa细胞周期进展和增殖。