Yang Botao, Wu Yucai, Chen Yang, Li Yongshuang, Wang Jinhua, Cha Xushan, Liu Jing
Department of Dermatology, The First Affiliated Clinical College of Guangzhou University of Chinese Medicine, Guangzhou, Guangdong Province, China.
Department of Dermatology, Guangdong Provincial People's Hospital's Nanhai Hospital, Foshan, Guangdong Province, China.
Heliyon. 2023 Aug 19;9(9):e19227. doi: 10.1016/j.heliyon.2023.e19227. eCollection 2023 Sep.
Although miR-5195-3p has been acknowledged for its tumor suppressor role in diverse cancer categories, its precise functions and mechanisms concerning melanoma have not been comprehensively elucidated. In this study, we employed quantitative reverse transcription PCR, Western blot analysis, and immunohistochemistry staining to investigate the expression patterns of miR-5195-3p and poly (rC) binding protein 2 (PCBP2) in melanoma tissues compared to adjacent tissues. Our findings revealed downregulation of miR-5195-3p and upregulation of PCBP2 in melanoma tissues. Through the implementation of a luciferase reporter assay, we successfully identified PCBP2 as a newly discovered target of miR-5195-3p in melanoma cells. Enforced expression of miR-5195-3p via mimics inhibited cell proliferation and migration in A375 and A2058 cells, as demonstrated by CCK-8 and transwell migration assays. In melanoma cells, reintroduction of PCBP2 partially reversed the inhibitory effects of miR-5195-3p overexpression. Treatment with LY294002, an inhibitor of the PI3K/AKT signaling pathway, also reversed the effects of PCBP2 in melanoma cells. Furthermore, our results suggest that miR-5195-3p inhibits the activation of the PI3K/AKT signaling pathway in melanoma by inhibiting PCBP2. In conclusion, our research has identified the miR-5195-3p targeting of the PCBP2-mediated PI3K/AKT signaling pathway as a potential therapeutic target for melanoma treatment.
尽管miR-5195-3p因其在多种癌症类型中的肿瘤抑制作用而得到认可,但其在黑色素瘤中的精确功能和机制尚未得到全面阐明。在本研究中,我们采用定量逆转录PCR、蛋白质免疫印迹分析和免疫组织化学染色,以研究miR-5195-3p和聚(rC)结合蛋白2(PCBP2)在黑色素瘤组织与相邻组织中的表达模式。我们的研究结果显示,黑色素瘤组织中miR-5195-3p表达下调,PCBP2表达上调。通过荧光素酶报告基因检测,我们成功确定PCBP2是黑色素瘤细胞中miR-5195-3p新发现的靶点。如CCK-8和Transwell迁移实验所示,通过模拟物强制表达miR-5195-3p可抑制A375和A2058细胞的增殖和迁移。在黑色素瘤细胞中,重新引入PCBP2可部分逆转miR-5195-3p过表达的抑制作用。用PI3K/AKT信号通路抑制剂LY294002处理也可逆转PCBP2在黑色素瘤细胞中的作用。此外,我们的结果表明,miR-5195-3p通过抑制PCBP2来抑制黑色素瘤中PI3K/AKT信号通路的激活。总之,我们的研究确定了miR-5195-3p靶向PCBP2介导的PI3K/AKT信号通路作为黑色素瘤治疗的潜在靶点。