Rui Zhao, Yanmin Yi, Han Liu, Jianwei Xu, Shuhai Chen, Dong Wu, Lei Wang, Feng Li
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引用次数: 0
Abstract
Background: The influence of the small Rho GTPase Rif (RHOF) on tumor growth, glycolysis, endothelial-mesenchymal transition (EMT), and the potential mechanism of RHOF in pancreatic cancer (PC) were explored.
Methods: RHOF expression in PC tissues and cells was assessed by qRT-PCR and western blotting. The viability, proliferation, apoptosis, migration, and invasion of PC cells were assessed using CCK-8, colony formation, EdU, flow cytometry, scratch, and Transwell assays. The expression of EMT- and glycolysis-related proteins was determined using western blotting. The potential mechanisms of action of RHOF in PC were identified using bioinformatic analysis. The effects of RHOF were assessed in vivo using a xenograft mouse model.
Results: PC cell proliferation, migration, and invasion are accelerated by RHOF overexpression, which inhibited apoptosis. RHOF overexpression promoted EMT and glycolysis as evidenced by a decrease in E-cadherin expression and an increase in N-cadherin, Vimentin, HK2, PKM2, and LDHA expression. Bioinformatic analysis indicated that RHOF activated EMT, glycolysis, and Myc targets and that c-Myc could bind to the PKM2 promoter. RHOF overexpression promotes the lactylation and nuclear translocation of Snail1. Silencing Snail1 reversed the promoting effects of RHOF and lactate on cell migration, invasion, and EMT. Moreover, in vivo tumor growth and EMT were inhibited by RHOF silencing.
Conclusion: RHOF plays an oncogenic role in PC. c-Myc is upregulated by RHOF and promotes PKM2 transcription. PKM2 further induces glycolysis, and the lactate produced by glycolysis causes the lactylation of Snail1, ultimately promoting EMT.
背景:探讨了小Rho GTP酶Rif(RHOF)对肿瘤生长、糖酵解、内皮-间质转化(EMT)的影响,以及RHOF在胰腺癌(PC)中的潜在机制:方法:采用 qRT-PCR 和免疫印迹法评估 RHOF 在 PC 组织和细胞中的表达。采用 CCK-8、集落形成、EdU、流式细胞术、划痕和 Transwell 试验评估了 PC 细胞的活力、增殖、凋亡、迁移和侵袭。采用 Western 印迹法测定了 EMT 和糖酵解相关蛋白的表达。利用生物信息学分析确定了 RHOF 在 PC 中的潜在作用机制。使用异种移植小鼠模型评估了RHOF在体内的作用:结果:过表达 RHOF 会加速 PC 细胞的增殖、迁移和侵袭,并抑制细胞凋亡。RHOF过表达促进了EMT和糖酵解,表现为E-cadherin表达减少,N-cadherin、Vimentin、HK2、PKM2和LDHA表达增加。生物信息学分析表明,RHOF激活了EMT、糖酵解和Myc靶标,而且c-Myc能与PKM2启动子结合。RHOF的过表达促进了Snail1的乳化和核转位。沉默Snail1可逆转RHOF和乳酸对细胞迁移、侵袭和EMT的促进作用。此外,RHOF沉默可抑制体内肿瘤生长和EMT:RHOF在PC中起着致癌作用。PKM2进一步诱导糖酵解,糖酵解产生的乳酸导致蜗牛1乳化,最终促进EMT。
期刊介绍:
Cancer & Metabolism welcomes studies on all aspects of the relationship between cancer and metabolism, including: -Molecular biology and genetics of cancer metabolism -Whole-body metabolism, including diabetes and obesity, in relation to cancer -Metabolomics in relation to cancer; -Metabolism-based imaging -Preclinical and clinical studies of metabolism-related cancer therapies.