Department of Transplantation, The Second Affiliated Hospital of Hainan Medical University, Haikou, 570100, China.
Department of Transplantation, The Hainan General Hospital, Haikou, 570100, China.
BMC Cancer. 2024 Apr 25;24(1):525. doi: 10.1186/s12885-024-12275-w.
Regorafenib, a multi-targeted kinase inhibitor, has been used in the treatment of Hepatocellular carcinoma (HCC). The purpose of this study is to investigate the mechanism of Regorafenib in HCC.
Regorafenib's impact on the sensitivity of HCC cells was assessed using CCK8. Differential gene expression analysis was performed by conducting mRNA sequencing after treatment with Regorafenib. The m6A methylation status of CHOP and differential expression of m6A methylation-related proteins were assessed by RIP and Western Blot. To explore the molecular mechanisms involved in the therapeutic effects of Regorafenib in HCC and the impact of METTL14 and CHOP on Regorafenib treatment, we employed shRNA/overexpression approaches to transfect METTL14 and CHOP genes, as well as conducted in vivo experiments.
Treatment with Regorafenib led to a notable decrease in viability and proliferation of SK-Hep-1 and HCC-LM3 cells. The expression level of CHOP was upregulated after Regorafenib intervention, and CHOP underwent m6A methylation. Among the m6A methylation-related proteins, METTL14 exhibited the most significant downregulation. Mechanistic studies revealed that Regorafenib regulated the cell cycle arrest in HCC through METTL14-mediated modulation of CHOP, and the METTL14/CHOP axis affected the sensitivity of HCC to Regorafenib. In vivo, CHOP enhanced the anticancer effect of Regorafenib.
The inhibition of HCC development by Regorafenib is attributed to its modulation of m6A expression of CHOP, mediated by METTL14, and the METTL14/CHOP axis enhances the sensitivity of HCC to Regorafenib. These findings provide insights into the treatment of HCC and the issue of drug resistance to Regorafenib.
瑞戈非尼是一种多靶点激酶抑制剂，已被用于治疗肝细胞癌（HCC）。本研究旨在探讨瑞戈非尼在 HCC 中的作用机制。
通过 CCK8 评估瑞戈非尼对 HCC 细胞敏感性的影响。用瑞戈非尼处理后进行 mRNA 测序，分析差异基因表达。通过 RIP 和 Western blot 评估 CHOP 的 m6A 甲基化状态和 m6A 甲基化相关蛋白的差异表达。通过 shRNA/过表达方法转染 METTL14 和 CHOP 基因，以及进行体内实验，探讨瑞戈非尼治疗 HCC 的疗效的分子机制以及 METTL14 和 CHOP 对瑞戈非尼治疗的影响。
瑞戈非尼治疗导致 SK-Hep-1 和 HCC-LM3 细胞活力和增殖明显下降。瑞戈非尼干预后 CHOP 的表达水平上调，CHOP 发生 m6A 甲基化。在 m6A 甲基化相关蛋白中，METTL14 的表达下调最为显著。机制研究表明，瑞戈非尼通过 METTL14 介导的 CHOP 调节来调控 HCC 中的细胞周期停滞，并且 METTL14/CHOP 轴影响 HCC 对瑞戈非尼的敏感性。在体内，CHOP 增强了瑞戈非尼的抗癌作用。
瑞戈非尼抑制 HCC 发展归因于其对 CHOP 的 m6A 表达的调节，这种调节是由 METTL14 介导的，而 METTL14/CHOP 轴增强了 HCC 对瑞戈非尼的敏感性。这些发现为 HCC 的治疗和瑞戈非尼耐药问题提供了新的思路。
