WTAP 介导的 circCMTM3 m6A 修饰通过招募 IGF2BP1 增加 PARK7 稳定性来抑制肝细胞癌铁死亡。
WTAP-mediated m6A modification on circCMTM3 inhibits hepatocellular carcinoma ferroptosis by recruiting IGF2BP1 to increase PARK7 stability.
机构信息
Department of Hepatobiliary Surgery, Chenzhou First People's Hospital of Hunan Province, Chenzhou, Hunan 423000, PR China.
Interventional Diagnosis and Treatment Center, Chenzhou First People's Hospital of Hunan Province, Chenzhou, Hunan 423000, PR China.
出版信息
Dig Liver Dis. 2023 Jul;55(7):967-981. doi: 10.1016/j.dld.2022.12.005. Epub 2022 Dec 30.
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, 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.
背景
肝细胞癌（HCC）预后不良，死亡率高。环状 RNA CMTM3 在 HCC 中显著上调。然而，circCMTM3 在 HCC 中的作用机制尚未完全阐明。
方法
通过 qRT-PCR、免疫组化和 Western blot 定量检测 HCC 细胞和组织中 circCMTM3、PARK7、GPX4 和 Ki67 的表达水平。试剂盒检测 GSH、总铁、Fe 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。MeRIP-PCR 评估 circCMTM3 的 m6A 水平。RNA 下拉、RIP 和 FISH 检测 circCMTM3、WTAP 和 PARK7 之间的相互作用。构建肿瘤异种移植模型以验证 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 的铁死亡。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 通过调节 PARK7 信号通路抑制裸鼠肿瘤生长并促进 HCC 铁死亡。
结论
circCMTM3 通过募集 IGF2BP1 增加 HCC 中的 PARK7 稳定性来抑制铁死亡，从而促进肝癌的发生，表明 cicrCMTM3 可能是 HCC 治疗的重要标志物。
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