Yao Xuming, Yang Zhiping, Hou Guoxin, Jiang Jialu, Wang Lvbin, Jiang Jin
Department of Oncology, The Affliated Hospital of Jiaxing University (The First Hospital of Jiaxing), Jiaxing, Zhejiang, China.
J Chemother. 2024 Sep 2:1-12. doi: 10.1080/1120009X.2024.2376422.
Colorectal cancer (CRC) is the second leading cause of cancer death, and about 10% of all malignancies are CRC. Cancer stem cells are considered main culprits in CRC treatment resistance and disease recurrence. This study explored the effects of tripartite motif containing 24 (TRIM24) and zinc finger protein, X-linked (ZFX) on CRC cell stemness and 5-FU resistance. A 5-FU-resistant cell line (HT29-5-FU) was constructed for functional analysis of CRC 5-FU-resistant cells. qRT-PCR and western blot (WB) were employed to analyze mRNA and protein levels of ZFX in 5-FU resistant cells and sensitive cells. WB was also utilized to analyze the surface markers of stem cells in each group. CCK-8 assay determined the IC values of different cell groups treated with 5-FU. The sphere-forming ability of cells in each group was determined using tumor sphere assay. Dual-luciferase reporter gene assay validated binding of ZFX to TRIM24. ZFX was highly expressed in HT29-5-FU cells. Silencing ZFX significantly reduced the 5-FU resistance and IC value of HT29-5-FU cells, and the surface markers and cell sphere-forming ability of stem cells were also significantly reduced. The function of HT29 cells was opposite when ZFX was overexpressed. In CRC cells, TRIM24 was an upstream transcription factor of ZFX, and they interacted with each other. TRIM24 activated the expression of ZFX to influence the stemness and 5-FU resistance of cells. The TRIM24/ZFX regulatory axis affected the stemness of CRC cells and their sensitivity to 5-FU, providing potential drug targets for novel therapeutic avenues for CRC.
结直肠癌（CRC）是癌症死亡的第二大主要原因，所有恶性肿瘤中约10%为CRC。癌症干细胞被认为是CRC治疗耐药性和疾病复发的主要罪魁祸首。本研究探讨了含三联基序蛋白24（TRIM24）和X连锁锌指蛋白（ZFX）对CRC细胞干性和5-氟尿嘧啶（5-FU）耐药性的影响。构建了5-FU耐药细胞系（HT29-5-FU）用于CRC 5-FU耐药细胞的功能分析。采用qRT-PCR和蛋白质免疫印迹法（WB）分析5-FU耐药细胞和敏感细胞中ZFX的mRNA和蛋白水平。WB还用于分析每组中干细胞的表面标志物。CCK-8法测定不同细胞组经5-FU处理后的半数抑制浓度（IC）值。采用肿瘤球形成试验测定每组细胞的成球能力。双荧光素酶报告基因试验验证ZFX与TRIM24的结合。ZFX在HT29-5-FU细胞中高表达。沉默ZFX可显著降低HT29-5-FU细胞的5-FU耐药性和IC值，同时干细胞的表面标志物和成球能力也显著降低。过表达ZFX时，HT29细胞的功能则相反。在CRC细胞中，TRIM24是ZFX的上游转录因子，二者相互作用。TRIM24激活ZFX的表达以影响细胞的干性和5-FU耐药性。TRIM24/ZFX调控轴影响CRC细胞的干性及其对5-FU的敏感性，为CRC新的治疗途径提供了潜在的药物靶点。
