NT5E inhibition suppresses the growth of sunitinib‐resistant cells and EMT course and AKT/GSK‐3β signaling pathway in renal cell cancer

• Published in: IUBMB life 2019-01, Vol.71 (1), p.113-124
• Main Authors: Peng, Dan, Hu, Zhiquan, Wei, Xian, Ke, Xinwen, Shen, Yuanqing, Zeng, Xing
• Format: Article
• Language: English
• Subjects: 5'-Nucleotidase - genetics; AKT protein; AKT/GSK‐3β; Animals; Carcinoma, Renal Cell - drug therapy; Carcinoma, Renal Cell - genetics; Carcinoma, Renal Cell - pathology; Cell growth; Cell migration; Cell Movement - drug effects; Cell proliferation; Cell Proliferation - drug effects; Cholecystokinin; Clear cell-type renal cell carcinoma; Drug Resistance, Neoplasm - drug effects; EMT; Epithelial-Mesenchymal Transition - drug effects; Glycogen Synthase Kinase 3 beta - genetics; GPI-Linked Proteins - genetics; Humans; Immunohistochemistry; Kidney cancer; Kinases; Mesenchyme; Mice; Neoplasm Invasiveness - genetics; Neoplasm Invasiveness - pathology; NT5E; Nucleotidase; Proto-Oncogene Proteins c-akt - genetics; renal cell cancer; Signal transduction; Signal Transduction - drug effects; sunitinib; Sunitinib - pharmacology; Wound healing
• ISSN: 1521-6543; 1521-6551
• DOI: 10.1002/iub.1942

We aimed to explore the mechanisms of sunitinib‐resistance in renal cell cancer (RCC) and provide new therapeutic evidence and biomarkers for RCC treatment using human clear‐cell renal cell carcinoma (ccRCC) cell line, 786‐O. Microarray analysis, quantitative real time PCR (qRT‐PCR), Western blot, and immunohistochemistry were used to detect Ecto‐5′‐nucleotidase (NT5E) expressions in sunitinib‐resistant tissues in RCC. 3‐(4,5‐dimethyl‐2‐thiazolyl)‐2,5‐diphenyl‐2‐H‐tetrazolium bromide, Thiazolyl Blue Tetrazolium Bromide (MTT), cell‐count kit 8 (CCK‐8), wound healing, and Transwell assays were used to investigate the influences of sunitinib and NT5E on 786‐O cell line and sunitinib‐resistant 786‐O cell line (S786‐O). Protein expressions related to epithelial–mesenchymal transition (EMT) and AKT/GSK‐3β signaling pathway in 786‐O cells and S786‐O cells were determined by Western blot. In vivo experiment was performed using NCr‐nu/nu mice to explore the effects of NT5E on cell growth and EMT signal in sunitinib environment. NT5E expression was upregulated in sunitinib‐resistant RCC tissues and cells. NT5E downregulation repressed RCC cell proliferation, migration as well as invasion. EMT course and AKT/GSK‐3β signal pathway both in vitro and in vivo in sunitinib environment was suppressed to varying degrees via NT5E inhibition. NT5E inhibition could suppress the growth of sunitinib‐resistant RCC cells and restrain EMT course and AKT/GSK‐3β signal pathway in sunitinib environment in RCC. © 2018 IUBMB Life, 71(1):113–124, 2019