Tianjin Key Laboratory of Urology, Department of Urology, The Second Hospital of Tianjin Medical University, Tianjin 300211, China.
Department of Urology, Beijing Aerospace General Hospital, Beijing, China.
Bioengineered. 2021 Dec;12(2):9377-9389. doi: 10.1080/21655979.2021.1974645.
Cartilage acidic protein 1 (CRTAC1) is predicted to be aberrantly expressed in bladder cancer based on bioinformatics analysis. However, its functions and molecular mechanism in bladder cancer remain elusive. This study aimed to explore the role of CRTAC1 in bladder cancer. The mRNA and protein levels of CRTAC1 and Yin Yang 1 (YY1) were detected by reverse transcription quantitative polymerase chain reaction and western blotting. We found that CRTAC1 was downregulated in bladder cancer tissues and cells. Cell Counting Kit-8 assays, colony formation assays, wound healing assays and Transwell assays and western blotting revealed that CRTAC1 overexpression inhibited cell viability, proliferation, migration, invasion and epithelial-mesenchymal transition (EMT) process in bladder cancer, while CRTAC1 knockdown exerted opposite effects on these malignant behaviors. Mechanistically, CRTAC1 targeted YY1 in bladder cancer cells. YY1 was upregulated in bladder cancer tissues and cells. CRTAC1 negatively modulated the mRNA and protein expression of YY1 in bladder cancer cells. Co-localization of CRTAC1 and YY1 expression was assessed using immunofluorescence staining and Co-Immunoprecipitation assays. The interaction between CRTAC1 and YY1 was explored by Chromatin immunoprecipitation and luciferase reporter assays. Moreover, CRTAC1 inactivated the TGF-β pathway by downregulating YY1 expression. Protein levels of factors associated with the TGF-β pathway were examined by western blotting. Rescue assays indicated that CRTAC1 inhibited malignant behaviors of bladder cancer cells by targeting YY1. Overall, CRTAC1 inhibited malignant phenotypes of bladder cancer cells by targeting YY1 to inactivate the TGF-β pathway.
基于生物信息学分析预测软骨酸性蛋白 1（CRTAC1）在膀胱癌中异常表达。然而，其在膀胱癌中的功能和分子机制仍不清楚。本研究旨在探讨 CRTAC1 在膀胱癌中的作用。采用逆转录定量聚合酶链反应和 Western blot 检测 CRTAC1 和 Yin Yang 1（YY1）的 mRNA 和蛋白水平。结果发现，CRTAC1 在膀胱癌组织和细胞中表达下调。细胞计数试剂盒-8 检测、集落形成实验、划痕愈合实验和 Transwell 实验以及 Western blot 显示，CRTAC1 过表达抑制膀胱癌细胞活力、增殖、迁移、侵袭和上皮-间充质转化（EMT）过程，而 CRTAC1 敲低则对这些恶性行为产生相反的影响。机制上，CRTAC1 在膀胱癌细胞中靶向 YY1。YY1 在膀胱癌组织和细胞中上调。CRTAC1 负调控膀胱癌细胞中 YY1 的 mRNA 和蛋白表达。采用免疫荧光染色和 Co-Immunoprecipitation 实验评估 CRTAC1 和 YY1 表达的共定位。采用 Chromatin immunoprecipitation 和 luciferase reporter 实验探索 CRTAC1 和 YY1 之间的相互作用。此外，CRTAC1 通过下调 YY1 表达来抑制 TGF-β 通路。采用 Western blot 检测与 TGF-β通路相关的因子的蛋白水平。通过挽救实验表明，CRTAC1 通过靶向 YY1 抑制膀胱癌细胞的恶性表型。总之，CRTAC1 通过靶向 YY1 抑制 TGF-β 通路抑制膀胱癌细胞的恶性表型。
