WTAP-mediated m6A modification on circCMTM3 inhibits hepatocellular carcinoma ferroptosis by recruiting IGF2BP1 to increase PARK7 stability
WTAP-mediated m6A modification on circCMTM3 inhibits hepatocellular carcinoma ferroptosis by recruiting IGF2BP1 to increase PARK7 stability
复制标题
DOI：
10.1016/j.dld.2022.12.005
复制
发表时间：
2023-06-26
影响因子：
4.5
通讯作者：
Sheng, Langqing
中科院分区：
文献类型：
--
作者：
Chen, Shuwei;Xia, Hongxiang;Sheng, Langqing
背景：肝细胞癌（HCC）预后差、死亡率高。 CircCMTM3 在 HCC 中显着上调。然而，circCMTM3在HCC中的作用机制尚未完全阐明。方法：通过 qRT-PCR、IHC 和 Western blotting 定量 HCC 细胞和组织中 circCMTM3、PARK7、GPX4 和 Ki67 的表达水平。他们的试剂盒检测了GSH、总铁、Fe 2 + 和MDA 的水平。 CCK-8和流式细胞术分析分别用于评估细胞增殖和脂质ROS水平。通过 MeRIP-PCR 评估 circCMTM3 的 m6A 水平。 RNA Pulldown、RIP 和 FISH 检测了 circCMTM3、WTAP 和 PARK7 之间的相互作用。构建肿瘤异种移植模型来验证cicrCMTM3和WTAP的功能。结果：CircCMTM3 和 WTAP 在 HCC 组织和细胞中增强。 WTAP 的敲低抑制了 circCMTM3 的 m6A 修饰，从而促进了 HCC 铁死亡。 circCMTM3沉默通过与HCC细胞中的IGF2BP1结合抑制PARK7的表达和稳定性，最终诱导铁死亡。体内研究表明，沉默 WTAP 和 circCMTM3 可通过调节 PARK7 信号传导抑制裸鼠肿瘤生长并促进 HCC 铁死亡。结论：CircCMTM3在HCC中通过招募IGF2BP1增加PARK7稳定性来抑制铁死亡，从而促进癌的发生，提示cicrCMTM3可能是HCC治疗的重要标志物。
Background: Hepatocellular carcinoma (HCC) has poor prognosis and high mortality. CircCMTM3 was significantly up-regulated in HCC. However, the mechanism of circCMTM3 in HCC is not full elucidated. Methods: The expression level of circCMTM3, PARK7, GPX4, and Ki67 in HCC cells and tissues were quantified by qRT-PCR, IHC, and Western blotting. The level of GSH, total iron, Fe 2 + , and MDA were detected by their kits. CCK-8 and flow cytometry analysis were used to evaluated cell proliferation and lipid ROS level, respectively. m6A level of circCMTM3 was assessed by MeRIP-PCR. RNA pulldown, RIP, and FISH detected the interaction between circCMTM3, WTAP, and PARK7. Tumor xenograft model was constructed to validate the function of cicrCMTM3 and WTAP. Results: CircCMTM3 and WTAP were enhanced in HCC tissues and cells. Knockdown of WTAP inhibited m6A modification of circCMTM3, which promoted HCC ferroptosis. circCMTM3 silencing suppressed the expression and stability of PARK7 through binding with IGF2BP1 in HCC cells, which finally induced ferroptosis. In vivo studies demonstrated that silencing WTAP and circCMTM3 suppressed tumor growth and promoted HCC ferroptosis in nude mice by regulating PARK7 signaling. Conclusion: CircCMTM3 promoted the carcinogenesis through inhibiting ferroptosis by recruiting IGF2BP1 to increase PARK7 stability in HCC, suggesting that cicrCMTM3 may be an important marker for HCC treatment.