CEBPD amplification and overexpression in urothelial carcinoma: a driver of tumor metastasis indicating adverse prognosis
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Yu-Hui Wang1,2, Wen-Jeng Wu3,4,5,6,7, Wei-Jan Wang2, Hsuan-Ying Huang8, Wei-Ming Li3,4,5, Bi-Wen Yeh4,5,7, Ting-Feng Wu9, Yow-Ling Shiue10, Jim Jinn-Chyuan Sheu10,*, Ju-Ming Wang2,*, Chien-Feng Li8,9,11,12,13,*
1 Department of Pathology, Chi-Mei Medical Center, Tainan, Taiwan
2 Institute of Bioinformatics and Biosignal Transduction, National Cheng Kung University, Tainan, Taiwan
3 Graduate Institute of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan
4 Department of Urology, School of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan
5 Department of Urology, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan
6 Department of Urology, Kaohsiung Municipal Hsiao-Kang Hospital, Kaohsiung, Taiwan
7 Center for Stem Cell Research, Kaohsiung Medical University, Kaohsiung, Taiwan.
8 Department of Pathology, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Kaohsiung, Taiwan
9 Department of Biotechnology, Southern Taiwan University of Science and Technology, Tainan, Taiwan
10 Institute of Biomedical Science, National Sun Yat-Sen University, Kaohsiung, Taiwan
11 National Institute of Cancer Research, National Health Research Institutes, Tainan, Taiwan
12 Institute of Clinical Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan
13 Department of Internal Medicine and Cancer Center, Kaohsiung Medical University Hospital, Kaohsiung Medical University, Kaohsiung, Taiwan
* These authors contributed equally as senior authors of this work
Chien-Feng Li, email:
Keywords: urothelial carcinoma, 8q, CEBPD, amplification, MMP2
Received: May 03, 2015 Accepted: August 11, 2015 Published: August 17, 2015
The molecular aberrations responsible for the progression of urothelial carcinoma (UC) remain largely obscure. To search candidate driver oncogenes in UC, we performed array-based genomic hybridization (aCGH) on 40 UBUC samples. Amplification of 8q11.21 was preferentially identified in patients who developed disease-specific death (53.8%) and distal metastasis (50.0%) but was barely detected in non-eventful cases (3.7% and 0%, respectively). In order to quantify the expression of candidate genes harbored in 8q11.21, laser-capture microdissection coupled with RT-PCR was performed on 32 of the 40 cases submitted to aCGH. With this, we identified CEBPD mRNA expression as most significantly associated with gains of 8q11.21, suggesting amplification-driven expression. By performing CEBPD-specific FISH and immunohistochemistry on 295 UBUCs, we confirmed CEBPD amplification (21.3%) and overexpression (29.8%) were strongly related to each other (p<0.001). Moreover, both were associated with adverse clinicopathologic features and worse outcomes. Furthermore, the clinical significance of CEBPD expression was also confirmed in an independent cohort comprised of 340 UCs from the upper urinary tract. Interestingly, CEBPD knockdown suppressed cell proliferation, migration and, most significantly, cell invasion ability in UC cells. The latter phenotype is attributed to downregulation of MMP2 as identified by RT2 Profiler PCR array. Moreover, expression of CEBPD significantly enhanced MMP2 expression and transcriptional activation by directly binding to its promoter region, as confirmed by promoter reporter assay and chromatin immunoprecipitation assay. Conclusively, CEBPD amplification is a mechanism driving increased mRNA and protein expression that confers aggressiveness in UC through MMP2-mediated cell invasiveness.
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