Zhao Rui, Yi Yanmin, Liu Han, Xu Jianwei, Chen Shuhai, Wu Dong, Wang Lei, Li Feng
Department of Hepatobiliary Surgery, Qilu Hospital of Shandong University, Jinan, Shandong, 250012, China.
Department of Pancreatic Surgery, General Surgery, Qi Lu Hospital, Cheeloo College of Medicine, Shandong University, 107 Wenhuaxi Road, Jinan, Shandong, 250012, China.
Cancer Metab. 2024 Oct 26;12(1):32. doi: 10.1186/s40170-024-00362-2.
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.
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.
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.
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.
探讨小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）中的潜在作用机制。
采用qRT-PCR和蛋白质免疫印迹法评估PC组织和细胞中RHOF的表达。使用CCK-8、集落形成、EdU、流式细胞术、划痕和Transwell实验评估PC细胞的活力、增殖、凋亡、迁移和侵袭。通过蛋白质免疫印迹法检测EMT和糖酵解相关蛋白的表达。利用生物信息学分析确定RHOF在PC中的潜在作用机制。使用异种移植小鼠模型在体内评估RHOF的作用效果。
RHOF过表达加速了PC细胞的增殖、迁移和侵袭，抑制了细胞凋亡。RHOF过表达促进了EMT和糖酵解，表现为E-钙黏蛋白表达降低，N-钙黏蛋白、波形蛋白、己糖激酶2（HK2）、丙酮酸激酶M2（PKM2）和乳酸脱氢酶A（LDHA）表达增加。生物信息学分析表明，RHOF激活了EMT、糖酵解和Myc靶点，且c-Myc可与PKM2启动子结合。RHOF过表达促进了Snail1的乳酸化和核转位。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中发挥致癌作用。RHOF上调c-Myc并促进PKM2转录。PKM2进一步诱导糖酵解，糖酵解产生的乳酸导致Snail1乳酸化，最终促进EMT。
