Department of Occupational and Environmental Health, Key Laboratory of Environmental Medicine Engineering, Ministry of Education, School of Public Health, Southeast University, Nanjing, Jiangsu, China.
Huzhou Center for Disease Control and Prevention, Huzhou, Zhejiang, China.
J Cell Physiol. 2020 Mar;235(3):2325-2335. doi: 10.1002/jcp.29139. Epub 2019 Sep 20.
The long intergenic noncoding RNA, regulator of reprogramming (linc-ROR) has been reported to participate in tumorigenesis, while its functions and fundamental mechanisms in esophageal squamous cell carcinoma (ESCC) remain unclear. In this study, gain-of-function assays showed that linc-ROR upregulation enhanced cell viability, promoted cell proliferation, and inhibited apoptosis. Mechanistically, the regulatory network of linc-ROR/miR-204-5p/MDM2 was established with bioinformatics analysis and online databases, then validated via dual-luciferase reporter assays, RNA immunoprecipitation assays in ESCC cells. Linc-ROR positively regulates the expression of MDM2 as a molecular sponge of miR-204-5p. Moreover, results of western blot and coimmunoprecipitation indicated that linc-ROR overexpression enhanced the ubiquitination level of p53, and its downstream apoptosis-related genes have showed higher bcl-2 expression, lower bax, and cleaved caspase-3 expressions, while miR-204-5p could counteract with this effect. Finally, small interfering RNAs tailored to linc-ROR were established to further evaluate its effects on ESCC comprehensively. In summary, this study revealed that linc-ROR modulated cell apoptosis and regulated p53 ubiquitination via targeting miR-204-5p/MDM2 axis, which provides a novel therapeutic insight into treatments for ESCC.
长链非编码 RNA，重编程调节因子（linc-ROR）已被报道参与肿瘤发生，但其在食管鳞状细胞癌（ESCC）中的功能和基本机制尚不清楚。在这项研究中，功能获得实验表明，linc-ROR 的上调增强了细胞活力，促进了细胞增殖，并抑制了细胞凋亡。通过生物信息学分析和在线数据库建立了 linc-ROR/miR-204-5p/MDM2 的调控网络，然后通过双荧光素酶报告基因实验、ESCC 细胞中的 RNA 免疫沉淀实验进行验证。linc-ROR 作为 miR-204-5p 的分子海绵，正向调节 MDM2 的表达。此外，western blot 和 co-immunoprecipitation 的结果表明，linc-ROR 过表达增强了 p53 的泛素化水平，其下游凋亡相关基因显示出更高的 bcl-2 表达、更低的 bax 和 cleaved caspase-3 表达，而 miR-204-5p 可以抵消这种作用。最后，设计了针对 linc-ROR 的小干扰 RNA 进一步全面评估其对 ESCC 的影响。总之，这项研究表明，linc-ROR 通过靶向 miR-204-5p/MDM2 轴调节细胞凋亡和调节 p53 泛素化，为 ESCC 的治疗提供了新的治疗思路。
