Xu Juan, Jiang Nan, Shi Haijuan, Zhao Shanshan, Yao Shuzhong, Shen Huimin
The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong 510080, P.R. China.
Int J Oncol. 2017 Apr;50(4):1383-1391. doi: 10.3892/ijo.2017.3915. Epub 2017 Mar 16.
MicroRNAs (miRNAs) play important roles in transcriptional regulation by targeting the 3'-UTR of target genes which participate in various biological processes. We aimed to investigate the potential role of miR-28-5p in the process of ovarian cancer development through regulating N4BP1. We found that the mRNA expression level of miR-28-5p was significantly increased in ovarian cancer tissues in comparison with adjacent ovarian tissues by qRT-PCR (P<0.0001). We established that miR‑28‑5p promoted the progression of ovarian cancer cell proliferation using colony forming assay and MTT assay. Wound healing assay and the migration and invasion assay showed that miR‑28‑5p accelerated the migration and invasion abilities of ovarian cancer cells. Simultaneously, we showed that miR‑28‑5p promoted ovarian cancer cell cycle, and inhibited apoptosis by flow cytometry in vitro. Furthermore, the results showed that miR‑28‑5p promoted the growth of ovarian tumor by tumor formation assay in vivo. The results of western blot analysis indicated that miR‑28‑5p promoted the protein expression level of F-actin. Western blot analysis also demonstrated that miR‑28‑5p promoted the progress of epithelial-mesenchymal transition (EMT) in ovarian carcinoma cells. In addition, we found that miR‑28‑5p downregulated N4BP1 mRNA and protein expression by qRT-PCR and western blot analysis in human ovarian cancer. Therefore, our study indicated that miR‑28‑5p promoted the progression of ovarian cancer cell cycle, proliferation, migration and invasion, inhibited apoptosis, and induced the process of EMT through inhibition of N4BP1 in vitro. Moreover, miR‑28‑5p promoted the growth of ovarian tumor in vivo.
微小RNA（miRNA）通过靶向参与各种生物学过程的靶基因的3'-非翻译区（3'-UTR）在转录调控中发挥重要作用。我们旨在通过调节N4BP1来研究miR-28-5p在卵巢癌发生过程中的潜在作用。通过qRT-PCR我们发现，与相邻卵巢组织相比，miR-28-5p在卵巢癌组织中的mRNA表达水平显著升高（P<0.0001）。我们通过集落形成试验和MTT试验确定miR-28-5p促进卵巢癌细胞增殖进程。伤口愈合试验以及迁移和侵袭试验表明，miR-28-5p加速了卵巢癌细胞的迁移和侵袭能力。同时，我们通过体外流式细胞术表明miR-28-5p促进卵巢癌细胞周期进程并抑制细胞凋亡。此外，体内肿瘤形成试验结果表明miR-28-5p促进卵巢肿瘤生长。蛋白质印迹分析结果表明，miR-28-5p促进F-肌动蛋白的蛋白表达水平。蛋白质印迹分析还证明，miR-28-5p促进卵巢癌细胞上皮-间质转化（EMT）进程。此外，通过qRT-PCR和蛋白质印迹分析我们发现，在人卵巢癌中miR-28-5p下调N4BP1的mRNA和蛋白表达。因此，我们的研究表明，miR-28-5p在体外通过抑制N4BP1促进卵巢癌细胞周期进程、增殖、迁移和侵袭，抑制细胞凋亡，并诱导EMT进程。此外，miR-28-5p在体内促进卵巢肿瘤生长。
