Linc-ROR regulates apoptosis in esophageal squamous cell carcinoma via modulation of p53 ubiquitination by targeting miR-204-5p/MDM2
Linc-ROR regulates apoptosis in esophageal squamous cell carcinoma via modulation of p53 ubiquitination by targeting miR-204-5p/MDM2
复制标题
Linc-ROR 通过靶向 miR-204-5p/MDM2 调节 p53 泛素化来调节食管鳞状细胞癌的细胞凋亡。
DOI：
10.1002/jcp.29139
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发表时间：
2020-03-01
影响因子：
5.6
通讯作者：
Liu, Ran
中科院分区：
文献类型：
--
作者：
Gao, Han;Wang, Tian;Liu, Ran
长基因间非编码RNA（linc-ROR）参与了食管鳞癌的发生，但其在食管鳞癌中的作用机制尚不清楚。在这项研究中，功能获得性测定表明，linc-ROR上调增强细胞活力，促进细胞增殖，并抑制凋亡。通过生物信息学分析和在线数据库建立linc-ROR/miR-204- 5 p/MDM 2的调控网络，并通过双荧光素酶报告基因检测、RNA免疫沉淀检测等方法验证其调控机制。Linc-ROR作为miR-204- 5 p的分子海绵正调控MDM 2的表达。Western blot和免疫共沉淀结果表明，linc-ROR过表达增强了p53的泛素化水平，其下游凋亡相关基因bcl-2表达增强，bax表达降低，caspase-3表达减少，而miR-204- 5 p可抵消这种作用。最后，建立针对linc-ROR的小干扰RNA，以进一步全面评价其对食管鳞癌的作用。综上所述，本研究揭示linc-ROR通过靶向miR-204- 5 p/MDM 2轴调控细胞凋亡和p53泛素化，为食管鳞癌的治疗提供了新的思路。
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.