Song Hongning, Lai Linfeng, Liu Min, Wang Xuxia, Zhang Jun, Zhang Shanyong
Department of Oral and Maxillofacial Surgery, School and Hospital of Stomatology, Shandong University and Shandong Provincial Key Laboratory of Oral Tissue Regeneration and Shandong Engineering Laboratory for Dental Materials and Oral Tissue Regeneration, Jinan, Shandong 250012, P.R. China.
Department of Stomatology, The Second Affiliated Hospital of Shandong First Medical University, Taian 271000, P.R. China.
Exp Ther Med. 2020 Jun;19(6):3707-3715. doi: 10.3892/etm.2020.8614. Epub 2020 Mar 19.
Oral squamous cell carcinoma (OSCC) is one of the most prevalent malignancies worldwide. MicroRNAs (miRNAs or miRs) serve crucial roles in the development of OSCC. miR-196a is upregulated in various tumors; however, the role of miR-196a in OSCC remains unclear. This present study aimed to determine the role and underlying mechanism of miR-196a in OSCC cells. Reverse transcription-quantitative PCR (RT-qPCR) was used to measure miR-196a levels in OSCC cells. MTT assays were also performed to determine cell proliferation. Cell migration was detected using wound healing assays and transwell assays, and cell apoptosis was analyzed via flow cytometry. The results indicated that the expression of miR-196a was increased in OSCC cells compared with normal oral squamous cells. TargetScan and luciferase reporter assays also confirmed that forkhead box O1 (FOXO1) was a target gene of miR-196a. It was demonstrated that FOXO1 small interfering RNA significantly promoted SCC9 cell proliferation and migration, and inhibited cell apoptosis. Furthermore, inhibition of miR-196a suppressed SCC9 cell proliferation and migration, and induced cell apoptosis. However, all effects of the miR-196a inhibitor were reversed following FOXO1 inhibition. Western blotting and RT-qPCR were subsequently performed to determine the effect of miR-196a on the PI3K/Akt signaling pathway. In the present study, transfection of miR-196a inhibitor suppressed the expression of phosphorylated (p)-PI3K and p-Akt, and enhanced the levels of FOXO1, while inhibition of FOXO1 exerted the opposite effects. Furthermore, it was demonstrated that miR-196a mimic significantly enhanced SCC9 cell proliferation and migration, and inhibited cell apoptosis. In conclusion, the results indicated that miR-196a serve as an oncogene in OSCCs. Downregulation of miR-196a inhibited the malignant biological processes of OSCC cells by targeting FOXO1. The current results may provide a novel therapeutic strategy for the treatment of patients with OSCC.
口腔鳞状细胞癌(OSCC)是全球最常见的恶性肿瘤之一。微小RNA(miRNA或miR)在OSCC的发生发展中发挥着关键作用。miR-196a在多种肿瘤中上调;然而,miR-196a在OSCC中的作用仍不清楚。本研究旨在确定miR-196a在OSCC细胞中的作用及其潜在机制。采用逆转录定量PCR(RT-qPCR)检测OSCC细胞中miR-196a的水平。还进行了MTT试验以测定细胞增殖。使用伤口愈合试验和Transwell试验检测细胞迁移,并通过流式细胞术分析细胞凋亡。结果表明,与正常口腔鳞状细胞相比,OSCC细胞中miR-196a的表达增加。TargetScan和荧光素酶报告基因试验也证实叉头框O1(FOXO1)是miR-196a的靶基因。结果表明,FOXO1小干扰RNA显著促进SCC9细胞增殖和迁移,并抑制细胞凋亡。此外,抑制miR-196a可抑制SCC9细胞增殖和迁移,并诱导细胞凋亡。然而,在抑制FOXO1后,miR-196a抑制剂的所有作用均被逆转。随后进行蛋白质印迹法和RT-qPCR以确定miR-196a对PI3K/Akt信号通路的影响。在本研究中,转染miR-196a抑制剂可抑制磷酸化(p)-PI3K和p-Akt的表达,并提高FOXO1的水平,而抑制FOXO1则产生相反的效果。此外,结果表明miR-196a模拟物显著增强SCC9细胞增殖和迁移,并抑制细胞凋亡。总之,结果表明miR-196a在OSCC中作为癌基因发挥作用。下调miR-196a通过靶向FOXO1抑制OSCC细胞的恶性生物学过程。目前的结果可能为OSCC患者的治疗提供一种新的治疗策略。