1. Academic Validation
  2. Wilms' tumor 1-associating protein promotes renal cell carcinoma proliferation by regulating CDK2 mRNA stability

Wilms' tumor 1-associating protein promotes renal cell carcinoma proliferation by regulating CDK2 mRNA stability

  • J Exp Clin Cancer Res. 2018 Feb 27;37(1):40. doi: 10.1186/s13046-018-0706-6.
Jingyuan Tang 1 2 Feng Wang 1 Gong Cheng 1 Shuhui Si 3 Xi Sun 4 Jie Han 1 Hao Yu 1 Wei Zhang 1 Qiang Lv 5 Ji-Fu Wei 6 Haiwei Yang 7
Affiliations

Affiliations

  • 1 Department of Urology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, 210029, China.
  • 2 Department of Urology, Jiangsu Province Hospital of TCM, Affiliated Hospital of Nanjing University of TCM, Nanjing, 210029, China.
  • 3 Research Division of Clinical Pharmacology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, 210029, China.
  • 4 Jiangsu Breast Disease Center, The First Affiliated Hospital of Nanjing Medical University, 300 Guangzhou Road, Nanjing, 210029, China.
  • 5 Department of Urology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, 210029, China. [email protected].
  • 6 Research Division of Clinical Pharmacology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, 210029, China. [email protected].
  • 7 Department of Urology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, 210029, China. [email protected].
Abstract

Background: Wilms' tumor 1-associating protein (WTAP) plays an important role in physiological processes and the development of tumor such as cell cycle regulation. The regulation of cell cycle is mainly dependent on cyclins and cyclin-dependent protein kinases (CDKs). Recent studies have shown that CDKs are closely related to the tumor diagnosis, progression and response to treatment. However, their specific biological roles and related mechanism in renal cell carcinoma (RCC) remain unknown.

Methods: Quantitative Real-Time PCR, western blotting and immunohistochemistry were used to detect the expression of WTAP and CDK2. The survival analysis was adopted to explore the association between WTAP expression and the prognosis of RCC. Cells were stably transfected with lentivirus approach and cell proliferation and cell cycle, as well as tumorigenesis in nude mice were performed to assess the effect of WTAP in RCC. RNA immunoprecipitation, Luciferase reporter assay and siRNA were employed to identify the direct binding sites of WTAP with CDK2 transcript. Colony formation assay was conducted to confirm the function of CDK2 in WTAP-induced growth promoting.

Results: In RCC cell lines and tissues, WTAP was significantly over-expressed. Compared with patients with low expression of WTAP, patients with high expression of WTAP had lower overall survival rate. Additionally, cell function test indicated that cell proliferation abilities in WTAP over-expressed group were enhanced, while WTAP knockdown showed the opposite results. Subcutaneous xenograft tumor model displayed that knockdown of WTAP could impede tumorigenesis in vivo. Mechanism study exhibited that CDK2 expression was positively associated with the expression of WTAP. Moreover, WTAP stabilized CDK2 transcript to enhance CDK2 expression via binding to 3'-UTR of CDK2 transcript. Additionally, specific inhibitors of CDK2 activity and small interfering RNA (siRNA) of CDK2 expression inhibited WTAP-mediated promotion of proliferation.

Conclusions: These findings suggest that WTAP may have an oncogenic role in RCC through physically binding to CDK2 transcript and enhancing its transcript stability which might provide new insights into RCC therapy.

Keywords

CDK2; Prognosis; Proliferation; Renal cell carcinoma; WTAP.

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