Zou Feng, Zhang Guofei, Mei Gang, Zhang Huantao, Xie Mengliang, Dan Mingjiang
Department of Urology, The Seventh Affiliated Hospital, Southern Medical University Foshan 528000, Guangdong, China.
Department of Orthopedics, The Seventh Affiliated Hospital, Southern Medical University Foshan 528000, Guangdong, China.
Am J Transl Res. 2024 Jul 15;16(7):3248-3258. doi: 10.62347/QWAK3951. eCollection 2024.
To investigate the role of C-terminal tensin-like (CTEN) in mediating chemotherapy resistance via epithelial-mesenchymal transition (EMT) in bladder cancer (BC) cells, through the regulation of transforming growth factor-β1 (TGF-β1) expression.
Lentiviral vectors were used to create CTEN overexpression and knockdown constructs, which were then introduced into paclitaxel-resistant BC cell lines. The effects of CTEN manipulation on cell proliferation and drug sensitivity was assessed using the CCK-8 assay, and apoptosis was evaluated by flow cytometry. The expression levels of CTEN, TGF-β1, and EMT markers were quantified by RT-qPCR and Western blot analysis. The interaction between CTEN and TGF-β1 and its effect on TGF-β1 methylation were studied using bisulfite sequencing PCR and co-immunoprecipitation.
Overexpression of CTEN in BC cells was associated with decreased paclitaxel efficacy, reduced apoptosis, and elevated levels of TGF-β1 and EMT-related proteins. CTEN was found to bind TGF-β1, inhibiting its methylation and thereby promoting TGF-β1 upregulation. This increase in TGF-β1 expression facilitated the EMT process and enhanced drug resistance in BC cells.
The induction of TGF-β1 expression by CTEN promotes EMT and increases chemotherapy resistance in BC cells. Targeting CTEN or the EMT pathway could improve chemosensitivity in treatment-resistant BC, suggesting a novel therapeutic strategy to enhance chemotherapy effectiveness.
通过调节转化生长因子-β1（TGF-β1）的表达，研究C末端张力蛋白样分子（CTEN）在介导膀胱癌（BC）细胞上皮-间质转化（EMT）从而产生化疗耐药中的作用。
使用慢病毒载体构建CTEN过表达和敲低构建体，然后将其导入耐紫杉醇的BC细胞系。使用CCK-8法评估CTEN操作对细胞增殖和药物敏感性的影响，并通过流式细胞术评估细胞凋亡。通过RT-qPCR和蛋白质免疫印迹分析定量CTEN、TGF-β1和EMT标志物的表达水平。使用亚硫酸氢盐测序PCR和免疫共沉淀研究CTEN与TGF-β1之间的相互作用及其对TGF-β1甲基化的影响。
BC细胞中CTEN的过表达与紫杉醇疗效降低、细胞凋亡减少以及TGF-β1和EMT相关蛋白水平升高有关。发现CTEN与TGF-β1结合，抑制其甲基化，从而促进TGF-β1上调。TGF-β1表达的增加促进了EMT过程并增强了BC细胞的耐药性。
CTEN诱导TGF-β1表达促进了EMT并增加了BC细胞的化疗耐药性。靶向CTEN或EMT途径可提高耐药BC的化疗敏感性，提示一种增强化疗效果的新治疗策略。
