Zhu Xiaomei, Sun Jianfang
Department of Pathology, Institute of Dermatology, Chinese Academy of Medical Sciences and Peking Union Medical College, Nanjing, 210042, China.
Department of Pathology, Institute of Dermatology, Chinese Academy of Medical Sciences and Peking Union Medical College, Nanjing, 210042, China.
Mol Cell Probes. 2020 Oct;53:101644. doi: 10.1016/j.mcp.2020.101644. Epub 2020 Aug 13.
To investigate the role of circHIPK3 in melanoma.
Bioinformatics analysis and experiments including RT-qPCR, Pearson's correlation analysis, luciferase reporter, Western blot, and RIP assays were applied to explore the function and mechanism of circHIPK3 in melanoma.
CircHIPK3 expression was strikingly upregulated while miR-215-5p was downregulated in melanoma tissues and cell lines. Pearson's correlation analysis unveiled circHIPK3 expression was positively correlated with Ki-67 (a marker of proliferation), which implied that circHIPK3 may play a vital role in the progression of melanoma. In mechanism, luciferase reporter and RIP assays validated that circHIPK3 was able to bind with miR-215-5p. Moreover, we confirmed that overexpression of circHIPK3 could facilitate cell proliferation and depress cell apoptosis in melanoma while overexpression of miR-215-5p exerted opposite effects. Besides, our findings indicated that miR-215-5p overexpression significantly reversed the circHIPK3 overexpressing-mediated promotive effect on cell proliferation and inhibitory effect on cell apoptosis. Furthermore, we found that miR-215-5p could directly target YY1. Upregulation of YY1 could notably offset the inhibitory effect of circHIPK3 downregulation on cell proliferation and the promotive effect on cell apoptosis.
Our study corroborated that circHIPK3 regulated melanoma cell behaviors via the miR-215-5p/YY1 axis, which might provide a novel insight for the treatment of melanoma patients.
研究环状HIPK3在黑色素瘤中的作用。
应用生物信息学分析以及包括逆转录定量聚合酶链反应（RT-qPCR）、皮尔逊相关性分析、荧光素酶报告基因检测、蛋白质免疫印迹法（Western blot）和RNA免疫沉淀（RIP）分析等实验，探讨环状HIPK3在黑色素瘤中的功能及机制。
在黑色素瘤组织和细胞系中，环状HIPK3表达显著上调，而miR-215-5p表达下调。皮尔逊相关性分析显示，环状HIPK3表达与Ki-67（一种增殖标志物）呈正相关，这表明环状HIPK3可能在黑色素瘤进展中起重要作用。机制上，荧光素酶报告基因检测和RIP分析证实环状HIPK3能够与miR-215-5p结合。此外，我们证实，环状HIPK3过表达可促进黑色素瘤细胞增殖并抑制细胞凋亡，而miR-215-5p过表达则产生相反作用。此外，我们的研究结果表明，miR-215-5p过表达显著逆转了环状HIPK3过表达介导的对细胞增殖的促进作用和对细胞凋亡的抑制作用。此外，我们发现miR-215-5p可直接靶向YY1。YY1上调可显著抵消环状HIPK3下调对细胞增殖的抑制作用和对细胞凋亡的促进作用。
我们的研究证实，环状HIPK3通过miR-215-5p/YY1轴调节黑色素瘤细胞行为，这可能为黑色素瘤患者的治疗提供新的见解。
