E2F1 靶向 ARHGEF39 促进脂肪酸代谢介导的肝细胞癌转移。
ARHGEF39 targeted by E2F1 fosters hepatocellular carcinoma metastasis by mediating fatty acid metabolism.
机构信息
Department of Hepatobiliary Surgery, The First Affiliated Hospital of Fujian Medical University, Fuzhou, Fujian Province, 350005, China; Department of Hepatobiliary Surgery, National Regional Medical Center, Binhai Campus of the First Affiliated Hospital, Fujian Medical University, Fuzhou, Fujian Province, 350212, China; Department of Hepatobiliary Surgery, Mengchao Hepatobiliary Hospital of Fujian Medical University, Fuzhou, Fujian Province, 350005, China.
Department of Hepatobiliary Surgery, Mengchao Hepatobiliary Hospital of Fujian Medical University, Fuzhou, Fujian Province, 350005, China.
出版信息
Clin Res Hepatol Gastroenterol. 2024 Oct;48(8):102446. doi: 10.1016/j.clinre.2024.102446. Epub 2024 Aug 9.
BACKGROUND
Hepatocellular carcinoma (HCC) stands as the prevailing manifestation of primary liver cancer. Previous studies have implicated ARHGEF39 in various cancer progression processes, but its impact on HCC metastasis remains unclear.
METHODS
Bioinformatics analysis and qRT-PCR were employed to test ARHGEF39 expression in HCC tissues and cells, identified enriched pathways associated with ARHGEF39, and investigated its regulatory relationship with E2F1. The impact of ARHGEF39 overexpression or knockdown on cellular phenotypes in HCC was assessed through the implementation of CCK-8 and Transwell assays. Accumulation of neutral lipids was determined by BODIPY 493/503 staining, while levels of triglycerides and phospholipids were measured using specific assay kits. Expression of E-cadherin, Vimentin, MMP-2, MMP-9, and FASN were analyzed by Western blot. The interaction between ARHGEF39 and E2F1 was validated through ChIP and dual-luciferase reporter assays.
RESULTS
Our study demonstrated upregulated expression of both ARHGEF39 and E2F1 in HCC, with ARHGEF39 being associated with fatty acid metabolism (FAM) pathways. Additionally, ARHGEF39 was identified as a downstream target gene of E2F1. Cell-based experiments unmasked that high expression of ARHGEF39 mediated the promotion of HCC cell viability, migration, and invasion via enhanced FAM. Moreover, rescue assays demonstrated that the promotion of HCC cell metastasis by high ARHGEF39 expression was attenuated upon treatment with Orlistat. Conversely, the knockdown of E2F1 suppressed HCC cell metastasis and FAM, while the upregulation of ARHGEF39 counteracted the repressive effects of E2F1 downregulation on the metastatic potential of HCC cells.
CONCLUSION
Our findings confirmed the critical role of ARHGEF39 in HCC metastasis and unmasked potential molecular mechanisms through which ARHGEF39 fostered HCC metastasis via FAM, providing a theoretical basis for exploring novel molecular markers and preventive strategies for HCC metastasis.
背景
肝细胞癌（HCC）是原发性肝癌的主要表现形式。先前的研究表明 ARHGEF39 参与了多种癌症进展过程，但它对 HCC 转移的影响尚不清楚。
方法
采用生物信息学分析和 qRT-PCR 检测 HCC 组织和细胞中 ARHGEF39 的表达，鉴定与 ARHGEF39 相关的富集途径，并研究其与 E2F1 的调控关系。通过 CCK-8 和 Transwell 测定评估 ARHGEF39 过表达或敲低对 HCC 细胞表型的影响。通过 BODIPY 493/503 染色测定中性脂质的积累，并用特定的测定试剂盒测定甘油三酯和磷脂的水平。通过 Western blot 分析 E-钙粘蛋白、波形蛋白、MMP-2、MMP-9 和 FASN 的表达。通过 ChIP 和双荧光素酶报告基因检测验证 ARHGEF39 与 E2F1 之间的相互作用。
结果
我们的研究表明，ARHGEF39 和 E2F1 在 HCC 中均上调表达，ARHGEF39 与脂肪酸代谢（FAM）途径有关。此外，ARHGEF39 被鉴定为 E2F1 的下游靶基因。细胞实验揭示，高表达的 ARHGEF39 通过增强 FAM 促进 HCC 细胞活力、迁移和侵袭。此外，挽救实验表明，用奥利司他处理可减弱高 ARHGEF39 表达对 HCC 细胞转移的促进作用。相反，E2F1 的敲低抑制 HCC 细胞转移和 FAM，而 ARHGEF39 的上调抵消了 E2F1 下调对 HCC 细胞转移潜能的抑制作用。
结论
我们的研究结果证实了 ARHGEF39 在 HCC 转移中的关键作用，并揭示了 ARHGEF39 通过 FAM 促进 HCC 转移的潜在分子机制，为探索 HCC 转移的新分子标志物和预防策略提供了理论依据。
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