Li Wei, Chen Jian, Yang Jinbao, Zhang Bo, Wen Dihao, Jiang Zhibin
General Surgery Department, The 980th Hospital of the People's Liberation Army Joint Logistics Support Force, No. 398, West Zhongshan Road, Qiaoxi District, Shijiazhuang City, Hebei Province, 050051, China.
Anticancer Agents Med Chem. 2025;25(15):1085-1093. doi: 10.2174/0118715206336851241204111721.
This study explored whether the cell cycle regulator cadherin 1 (CDH1) impacts colorectal cancer cell cycle and stemness via mediating ubiquitination of sirtuin 5 (SIRT5).
We first constructed CDH1 overexpression plasmid and small interfering RNA against SIRT5 (siSIRT5) and transfected them into HCT116/HT29 cells, followed by transfection efficiency verification. The effect of CDH1 on Cyclin F/SIRT5/CDH1 protein levels in HCT116/HT29 cells was verified by Western blot. After up-regulation of CDH1, changes in SIRT5 ubiquitination (immunoprecipitation), cell cycle (cell cycle kit), proliferation (5-Bromodeoxyuridine assay), and stemness marker expressions (qRT-PCR) in HCT116/HT29 cells were detected. Rescue assays were performed to examine cell proliferation and stemness marker expressions.
Overexpression of CDH1 decreased Cyclin F expression and increased SIRT5 and CDH1 expressions in HCT116/HT29 cells. Up-regulation of CDH1 suppressed SIRT5 ubiquitination, promoted G0/G1 phase blockage in HCT116/HT29 cells, boosted cell proliferation into quiescence and enhanced cell stemness. siSIRT5 counteracted the regulatory effect of CDH1 overexpression on colorectal cancer cells.
CDH1 promotes the entry of colorectal cancer cells into quiescence and enhances stemness by dampening SIRT5 ubiquitination.
本研究探讨细胞周期调节因子钙黏蛋白1（CDH1）是否通过介导沉默调节蛋白5（SIRT5）的泛素化影响结直肠癌细胞周期和干性。
我们首先构建了CDH1过表达质粒和针对SIRT5的小干扰RNA（siSIRT5），并将它们转染到HCT116/HT29细胞中，随后验证转染效率。通过蛋白质免疫印迹法验证CDH1对HCT116/HT29细胞中细胞周期蛋白F/SIRT5/CDH1蛋白水平的影响。上调CDH1后，检测HCT116/HT29细胞中SIRT5泛素化（免疫沉淀）、细胞周期（细胞周期试剂盒）、增殖（5-溴脱氧尿苷检测）和干性标志物表达（qRT-PCR）的变化。进行挽救实验以检测细胞增殖和干性标志物表达。
CDH1过表达降低了HCT116/HT29细胞中细胞周期蛋白F的表达，并增加了SIRT5和CDH1的表达。上调CDH1可抑制SIRT5泛素化，促进HCT116/HT29细胞的G0/G1期阻滞，使细胞增殖进入静止期并增强细胞干性。siSIRT5抵消了CDH1过表达对结直肠癌细胞的调节作用。
CDH1通过抑制SIRT5泛素化促进结直肠癌细胞进入静止期并增强干性。
