背景:肝细胞癌(HCC)是原发性肝癌的主要表现。以前的研究表明ARHGEF39参与了各种癌症进展过程,但其对HCC转移的影响尚不清楚。
方法:生物信息学分析和qRT-PCR检测ARHGEF39在HCC组织和细胞中的表达,确定了与ARHGEF39相关的富集途径,并研究了其与E2F1的调控关系。通过实施CCK-8和Transwell测定来评估ARHGEF39过表达或敲低对HCC中细胞表型的影响。通过BODIPY493/503染色确定中性脂质的积累,而甘油三酯和磷脂的水平使用特定的测定试剂盒测量。E-cadherin的表达,Vimentin,通过蛋白质印迹分析MMP-2、MMP-9和FASN。ARHGEF39和E2F1之间的相互作用通过ChIP和双荧光素酶报告基因测定进行了验证。
结果:我们的研究表明,在HCC中ARHGEF39和E2F1的表达上调,ARHGEF39与脂肪酸代谢(FAM)途径相关。此外,ARHGEF39被鉴定为E2F1的下游靶基因。基于细胞的实验揭示了高表达ARHGEF39介导的肝癌细胞活力的促进,迁移,并通过增强的FAM进行入侵。此外,抢救试验表明,ARHGEF39高表达对肝癌细胞转移的促进作用在奥利司他治疗后减弱.相反,E2F1的敲除抑制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:我们的发现证实了ARHGEF39在HCC转移中的关键作用,以及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理论依据。
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.
CONCLUSIONS: 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.