Department of Clinical Laboratory, The Second Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong, China.
Department of Healthy Examination, The Second Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong, China.
Dis Markers. 2022 Oct 25;2022:6138941. doi: 10.1155/2022/6138941. eCollection 2022.
The levels of MCF2L were detected by PCR and western blotting assay. The effect of MCF2L on ferroptosis was confirmed by MTT, colony formation assay, Brdu, in vivo animal experiment, and the content of Iron, GSH, ROS, and MDA. The underlying mechanisms were explored by PCR, western blotting, and affinity precipitation assay. Our findings demonstrated that MCF2L is remarkedly upregulated in HCC tissues, and sorafenib can induce the levels of MCF2L, suggesting that MCF2L might function in sorafenib resistance of HCC. Further analysis showed that downregulation of MCF2L enhances HCC cell death induced by sorafenib, and ferroptosis inhibitor can reverse this process. Subsequent experiments showed that downregulation of MCF2L elevates the content of Iron, ROS, and MDA, which are all indicators of ferroptosis. Finally, mechanism analysis showed that MCF2L regulates the PI3K/AKT pathway in a RhoA/Rac1 dependent manner.
Our study showed that targeting MCF2L may be a hopeful method to overcome sorafenib-resistance through inducing ferroptosis in HCC.
通过 PCR 和 Western blot 检测 MCF2L 的水平。通过 MTT、集落形成实验、Brdu、体内动物实验和铁、GSH、ROS 和 MDA 的含量来验证 MCF2L 对铁死亡的影响。通过 PCR、Western blot 和亲和沉淀实验探索潜在机制。我们的研究结果表明，MCF2L 在 HCC 组织中显著上调，索拉非尼可以诱导 MCF2L 的水平，这表明 MCF2L 可能在 HCC 对索拉非尼的耐药性中发挥作用。进一步的分析表明，下调 MCF2L 增强了索拉非尼诱导的 HCC 细胞死亡，铁死亡抑制剂可以逆转这一过程。随后的实验表明，下调 MCF2L 会升高铁、ROS 和 MDA 的含量，这些都是铁死亡的指标。最后，机制分析表明 MCF2L 通过 RhoA/Rac1 依赖的方式调节 PI3K/AKT 通路。
我们的研究表明，通过诱导 HCC 中的铁死亡来靶向 MCF2L 可能是克服索拉非尼耐药的一种有希望的方法。
