Tian Jing, Shen Rui, Yan Yuzhang, Deng Liehua
Department of Dermatology, The First Affiliated Hospital of Jinan University, Guangzhou, Guangdong 510632, P.R. China.
Department of Plastic Cosmetic Surgery, The First People's Hospital of Foshan, Foshan, Guangdong 528000, P.R. China.
Exp Ther Med. 2018 Nov;16(5):4010-4018. doi: 10.3892/etm.2018.6679. Epub 2018 Sep 3.
Cutaneous squamous cell carcinoma (cSCC) accounts for 20% of non-melanoma skin cancer worldwide. MicroRNAs (miRNAs or miRs) are a subtype of non-coding RNA associated with the progression of various types of human cancer. MiR-186 has been demonstrated to act as an oncogene in human tumors. However, the role of miR-186 in cSCC remains unclear. The expression of miR-186 and apoptotic protease activating factor 1 (APAF1) was examined using reverse transcription-quantitative polymerase chain reaction, western blotting and immunofluorescence. The correlation between miR-186 and APAF1 was determined using a dual-luciferase assay. Mimics or inhibitors of miR-186 were transfected into A-431 cells to establish cell lines with overexpressed or knocked-down miR-186, respectively. EdU staining and colony formation assays were performed to detect cell proliferation. Transwell and wound-healing assays were performed to analyze cell invasion and migration, respectively. Hoechst staining and flow cytometry were performed to assess cell apoptosis and cell cycle distribution. MiR-186 expression was significantly increased, while APAF1 expression was significantly decreased in cSCC tissues compared with the controls. An miR-186 binding site was predicted in APAF1 and their expression was negatively correlated in cSCC tissues. Cell proliferation, invasion and migration were significantly enhanced in the miR-186-overexpressed A-431 cells and attenuated in miR-186 knockdown cells compared with the control. APAF1 expression was regulated by miR-186, while APAF1 knockdown significantly promoted cell invasion and inhibited cell apoptosis. In summary, the results of the present study indicate that miR-186 serves as an oncogene in cSCC by inhibiting APAF1.
皮肤鳞状细胞癌(cSCC)占全球非黑色素瘤皮肤癌的20%。微小RNA(miRNA或miR)是一种非编码RNA亚型,与多种人类癌症的进展相关。已证明miR-186在人类肿瘤中起癌基因作用。然而,miR-186在cSCC中的作用仍不清楚。使用逆转录-定量聚合酶链反应、蛋白质印迹法和免疫荧光法检测miR-186和凋亡蛋白酶激活因子1(APAF1)的表达。使用双荧光素酶测定法确定miR-186与APAF1之间的相关性。将miR-186的模拟物或抑制剂转染到A-431细胞中,分别建立miR-186过表达或敲低的细胞系。进行EdU染色和集落形成试验以检测细胞增殖。分别进行Transwell试验和伤口愈合试验以分析细胞侵袭和迁移。进行Hoechst染色和流式细胞术以评估细胞凋亡和细胞周期分布。与对照组相比,cSCC组织中miR-186表达显著增加,而APAF1表达显著降低。在APAF1中预测到一个miR-186结合位点,且它们在cSCC组织中的表达呈负相关。与对照组相比,miR-186过表达的A-431细胞中细胞增殖、侵袭和迁移显著增强,而miR-186敲低细胞中则减弱。APAF1表达受miR-186调控,而APAF1敲低显著促进细胞侵袭并抑制细胞凋亡。总之,本研究结果表明,miR-186通过抑制APAF1在cSCC中起癌基因作用。