CDH1-involved Ubiquitination of SIRT5 Promotes the Entry of Colorectal Cancer Cells into Quiescence and Enhances Cell Stemness.

IF 3 4区医学 Q3 CHEMISTRY, MEDICINAL Anti-cancer agents in medicinal chemistry Pub Date : 2025-01-01 DOI:10.2174/0118715206336851241204111721

Wei Li, Jian Chen, Jinbao Yang, Bo Zhang, Dihao Wen, Zhibin Jiang

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Abstract

Background: 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).

Methods: 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.

Results: 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.

Conclusion: CDH1 promotes the entry of colorectal cancer cells into quiescence and enhances stemness by dampening SIRT5 ubiquitination.

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cdh1介导的SIRT5泛素化促进结直肠癌细胞进入静止状态并增强细胞干性

背景：本研究探讨细胞周期调节因子cadherin 1 （CDH1）是否通过介导sirtuin 5 （SIRT5）的泛素化影响结直肠癌细胞周期和干细胞性。方法：首先构建CDH1过表达质粒和SIRT5小干扰RNA (siSIRT5)，转染至HCT116/HT29细胞，验证转染效率。Western blot验证CDH1对HCT116/HT29细胞Cyclin 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-溴脱氧尿苷法）和干性标志物表达（qRT-PCR）的变化。通过挽救实验检测细胞增殖和干细胞标记物的表达。结果：过表达CDH1可降低HCT116/HT29细胞中Cyclin F的表达，升高SIRT5和CDH1的表达。上调CDH1抑制SIRT5泛素化，促进HCT116/HT29细胞G0/G1期阻滞，促进细胞增殖进入静止状态，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。结论：CDH1通过抑制SIRT5泛素化，促进结直肠癌细胞进入静止状态，增强细胞干性。

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来源期刊

Anti-cancer agents in medicinal chemistry ONCOLOGY-CHEMISTRY, MEDICINAL

CiteScore

5.10

自引率

3.60%

发文量

323

审稿时长

4-8 weeks

期刊介绍： Formerly: Current Medicinal Chemistry - Anti-Cancer Agents. Anti-Cancer Agents in Medicinal Chemistry aims to cover all the latest and outstanding developments in medicinal chemistry and rational drug design for the discovery of anti-cancer agents. Each issue contains a series of timely in-depth reviews and guest edited issues written by leaders in the field covering a range of current topics in cancer medicinal chemistry. The journal only considers high quality research papers for publication. Anti-Cancer Agents in Medicinal Chemistry is an essential journal for every medicinal chemist who wishes to be kept informed and up-to-date with the latest and most important developments in cancer drug discovery.