Zhu Liangming, Zhang Lei, Tang Ying, Zhang Fang, Wan Chao, Xu Liang, Guo Ping
Department of Stomatology, Yijishan Hospital, The First Affiliated Hospital of Wannan Medical College, Wuhu, Anhui 241000, P.R. China.
Jiangcheng Dental Clinic, Wuhu, Anhui 241000, P.R. China.
Exp Ther Med. 2021 Jun;21(6):549. doi: 10.3892/etm.2021.9981. Epub 2021 Mar 24.
The aim of the present study was to evaluate the expression levels of microRNA (miR)-363-3p and its underlying physiological function in oral squamous cell carcinoma (OSCC). miR-363-3p expression levels were measured in OSCC cell lines using reverse transcription-quantitative PCR. The role of miR-363-3p in OSCC cells was examined using gain-of-function assays . Cell proliferation was assessed using Cell Counting Kit-8, 5-ethynyl-2'-deoxyuridine assays and flow cytometry. Cell migration and invasion were evaluated in wound-healing and Transwell Matrigel assays. In addition, bioinformatics analysis predicted binding sites of miR-363-3p on sperm-specific antigen 2 (SSFA2). Luciferase reporter and RNA pull-down assays were conducted to test whether miR-363-3p interacted with SSFA2. miR-363-3p expression was downregulated in OSCC cell lines compared with that in the normal epithelial cell line (NHOK). Additionally, miR-363-3p overexpression suppressed OSCC cell proliferation, migration and invasion . SSFA2 was verified as a direct target of miR-363-3p, and SSFA2 overexpression partially counteracted the inhibitory effects of miR-363-3p on cell proliferation, migration and invasion in OSCC cell lines. Thus, miR-363-3p may serve as a tumor suppressor via targeting SSFA2 and may represent a potential therapeutic target for OSCC.
本研究的目的是评估微小RNA(miR)-363-3p在口腔鳞状细胞癌(OSCC)中的表达水平及其潜在的生理功能。使用逆转录定量PCR检测OSCC细胞系中miR-363-3p的表达水平。使用功能获得性试验检测miR-363-3p在OSCC细胞中的作用。使用细胞计数试剂盒-8、5-乙炔基-2'-脱氧尿苷试验和流式细胞术评估细胞增殖。在伤口愈合试验和Transwell基质胶试验中评估细胞迁移和侵袭。此外,生物信息学分析预测了miR-363-3p在精子特异性抗原2(SSFA2)上的结合位点。进行荧光素酶报告基因试验和RNA下拉试验以检测miR-363-3p是否与SSFA2相互作用。与正常上皮细胞系(NHOK)相比,OSCC细胞系中miR-363-3p的表达下调。此外,miR-363-3p过表达抑制了OSCC细胞的增殖、迁移和侵袭。SSFA2被证实为miR-363-3p的直接靶点,并且SSFA2过表达部分抵消了miR-363-3p对OSCC细胞系中细胞增殖、迁移和侵袭的抑制作用。因此,miR-363-3p可能通过靶向SSFA2发挥肿瘤抑制作用,并且可能代表OSCC的潜在治疗靶点。