2. Academic Validation
  3. ROC1 promotes the malignant progression of bladder cancer by regulating p-IκBα/NF-κB signaling

ROC1 promotes the malignant progression of bladder cancer by regulating p-IκBα/NF-κB signaling

  • J Exp Clin Cancer Res. 2021 May 7;40(1):158. doi: 10.1186/s13046-021-01935-5.
Qi Wu 1,  Xiaoqing Zhou 1,  Peng Li 1,  Mao Ding 2,  Shengjie You 1,  Zhaoyu Xu 1,  Junjie Ye 1,  Xuedong Chen 1,  Mingyue Tan 3,  Jun Wang 3,  Wei Wang 4,  Jianxin Qiu 5
Affiliations

Affiliations

  • 1 Department of Urology, The Sixth Affiliated Hospital of Wenzhou Medical University (The People's Hospital of Lishui), Zhejiang, 323000, China.
  • 2 Department of Nephrology, Renmin Hospital of Wuhan University, Wuhan, 430000, China.
  • 3 Department of Urology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, No.100 Haining Road, Hongkou District, Shanghai, 200080, China.
  • 4 Department of Urology, The First people's Hospital of Yancheng (Yancheng First Hospital, Affiliated Hospital of Nanjing University Medical School), No.66 South Renmin Road, Yancheng, 224000, Jiangsu, China. rulerwong@163.com.
  • 5 Department of Urology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, No.100 Haining Road, Hongkou District, Shanghai, 200080, China. jasonqiu@sjtu.edu.cn.
PMID: 33962660 DOI: 10.1186/s13046-021-01935-5
Abstract

Background: Regulator of cullins 1 (ROC1) is an important catalytic subunit of cullin-RING E3 Ligase. Nuclear factor-kappa B (NF-κB) signaling is closely related to tumor invasion and metastasis. Earlier, we reported that ROC1 was associated with a poor prognosis in patients with bladder Cancer (BCa). However, it is unclear whether ROC1 is involved in the NF-κB signaling associated with malignant BCa progression.

Methods: The expression of ROC1 and p65 in bladder Cancer and paracancerous tissues were detected by immunohistochemistry (IHC). Pearson correlation was used to assess correlation between ROC1 and p65 protein expressions. The wound-healing and transwell assays were used to monitor cell invasion and migration. The effect of ROC1 on the expression of key proteins in the NF-κB signaling was determined by immunofluorescence and western blot (WB). Cycloheximide (CHX), MG132 and immunoprecipitation assays were used to evaluate the effect of ROC1 on the ubiquitination of phosphorylated inhibitor of kappa B alpha (p-IκBα). A lung metastasis mouse model was generated to detect the role of ROC1 in tumor metastasis.

Results: We found that ROC1 was up-regulated in BCa tissues and cell lines, and high ROC1 levels were positively correlated with higher tumour grade, lymph node metastasis, distant metastasis and poor prognosis. Linear-regression analysis showed significant a Pearson correlation between ROC1 and nuclear p65 expression in BCa tissue microarray (TMA) samples. Functional studies demonstrated that ROC1 promoted BCa cell invasion and migration. In vitro and in vivo experiments showed that ROC1 activated NF-κB signaling by enhancing the ubiquitination of p-IκBα, which caused p65 nuclear translocation and promoted the transcription of some metastasis-related target genes, such as urokinase-type plasminogen activator receptor (uPAR), intracellular adhesion molecule 1 (ICAM1), vascular cell adhesion molecule 1 (VCAM1), and matrix metalloproteinase 9 (MMP9), resulting in promoting BCa metastasis.

Conclusion: ROC1 plays an important role in the progression of BCa and serves as a potential diagnostic and therapeutic target for patients with BCa.

Keywords

Bladder cancer; Metastasis; NF-κB; ROC1; Ubiquitination.

Figures
Products