- 作者: Yi-Chieh Yang, Yung-Wei Lin, Wei-Jiunn Lee, Feng-Ru Lai, Kuo-Hao Ho, Chih-Ying Chu, Kuo-Tai Hua, Ji-Qing Chen, Min-Che Tung, Michael Hsiao, Yu-Ching Wen & Ming-Hsien Chien
- 作者服務機構: 1.Department of Cancer Biology, Geisel School of Medicine at Dartmouth, Lebanon, NH, USA 2.Department of Medical Education and Research, Wan Fang Hospital, Taipei Medical University, Taipei, Taiwan 3.Department of Medical Research, Tungs’ Taichung MetroHarbor Hospital, Taichung, Taiwan 4.Department of Surgery, Tungs’ Taichung Metro Harbor Hospital, Taichung, Taiwan 5.Department of Urology, School of Medicine, College of Medicine and TMU Research Center of Urology and Kidney (TMU-RCUK), Taipei Medical University, Taipei, Taiwan 6.Department of Urology, Wan Fang Hospital, Taipei Medical University, 111, Section 3, Hsing Long Road, Taipei, 11696, Taiwan 7.Genomics Research Center, Academia Sinica, Taipei, Taiwan 8.Graduate Institute of Clinical Medicine, College of Medicine, Taipei Medical University, 250 Wu Hsing Street, Taipei, 11031, Taiwan 9.Graduate Institute of Toxicology, College of Medicine, National Taiwan University, Taipei, Taiwan 10.International Master/PhD Program in Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan 11.Pulmonary Research Center, Wan Fang Hospital, Taipei Medical University, Taipei, Taiwan 12.TMU Research Center of Cancer Translational Medicine, Taipei Medical University, Taipei, Taiwan 13.Traditional Herbal Medicine Research Center, Taipei Medical University Hospital, Taipei, Taiwan
- 中文摘要:
- 英文摘要:
Background
KH-type splicing regulatory protein (KHSRP, also called KSRP), a versatile RNA-binding protein, plays a critical role in various physiological and pathological conditions through modulating gene expressions at multiple levels. However, the role of KSRP in clear cell renal cell carcinoma (ccRCC) remains poorly understood.
Methods
KSRP expression was detected by a ccRCC tissue microarray and evaluated by an in silico analysis. Cell loss-of-function and gain-of-function, colony-formation, anoikis, and transwell assays, and an orthotopic bioluminescent xenograft model were conducted to determine the functional role of KRSP in ccRCC progression. Micro (mi)RNA and complementary (c)DNA microarrays were used to identify downstream targets of KSRP. Western blotting, quantitative real-time polymerase chain reaction, and promoter- and 3-untranslated region (3'UTR)-luciferase reporter assays were employed to validate the underlying mechanisms of KSRP which aggravate progression of ccRCC.
Results
Our results showed that dysregulated high levels of KSRP were correlated with advanced clinical stages, larger tumor sizes, recurrence, and poor prognoses of ccRCC. Neural precursor cell-expressed developmentally downregulated 4 like (NEDD4L) was identified as a novel target of KSRP, which can reverse the protumorigenic and prometastatic characteristics as well as epithelial-mesenchymal transition (EMT) promotion by KSRP in vitro and in vivo. Molecular studies revealed that KSRP can decrease NEDD4L messenger (m)RNA stability via inducing mir-629-5p upregulation and directly targeting the AU-rich elements (AREs) of the 3’UTR. Moreover, KSRP was shown to transcriptionally suppress NEDD4L via inducing the transcriptional repressor, Wilm's tumor 1 (WT1). In the clinic, ccRCC samples revealed a positive correlation between KSRP and mesenchymal-related genes, and patients expressing high KSRP and low NEDD4L had the worst prognoses.
Conclusion
The current findings unveil novel mechanisms of KSRP which promote malignant progression of ccRCC through transcriptional inhibition and post-transcriptional destabilization of NEDD4L transcripts. Targeting KSRP and its pathways may be a novel pharmaceutical intervention for ccRCC. - 中文關鍵字:
- 英文關鍵字: ccRCC, Progression, KSRP, NEDD4L, EMT