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Bendoraite A , Knouf EC , Garg KS , Parkin RK , Kroh EM , O'Briant KC , Ventura AP , Godwin AK , Karlan BY , Drescher CW , Urban N , Knudsen BS , Tewari M
Regulation of miR-200 family microRNAs and ZEB transcription factors in ovarian cancer: Evidence supporting a mesothelial-to-epithelial transition
Gynecologic Oncology. 2010 Jan;116(1) :117-125
PMID: ISI:000273108700022   
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Abstract
Objective. Our objective was to characterize the expression and function of the miR-200 family of microRNAs (miRNA) in ovarian carcinogenesis. Methods. We used qRT-PCR to examine expression of the miR-200 miRNA family and its predicted targets, the ZEB1 and ZEB2 transcriptional repressors, in primary cultures of normal cells from the surface of the ovary and in a panel of 70 ovarian cancer tissues and 15 ovarian cancer cell lines. We Studied the mechanisms of regulation of miR-200 miRNAs and ZEB transcription factors in ovarian cells using 3' UTR luciferase reporters, promoter luciferase reporters and siRNAs. Results. miR-200 family members are expressed at low or negligible levels in normal ovarian surface cells and substantially increase in expression in ovarian cancer, whereas expression of ZEB1 and ZEB2 shows the opposite pattern. There is reciprocal repression between nnR-200 family members and ZEB transcription factors, creating a double negative regulatory feedback loop resembling that reported in other cancer cell types. In contrast to epithelial cells from other sites, expression levels of miR-200 miRNAs and ZEB1/2 in cells from the Ovarian Surface are more consistent with a mesenchymal cell phenotype, potentially reflecting the mesothelial origin of the ovarian Surface. Conclusion. Analysis of ovarian cancer tissues suggests that ovarian surface cells acquire a more epithelial miR-200-ZEB1/2 phenotype as they undergo transformation, switching from a miR-200 familyLOW and ZEB1/2HIGH state to a miR-200 familyHIGH and ZEB1/2LOW phenotype. Collectively, our data support the mesothelial-to-epithelial (Meso-E-T) model for development of ovarian cancers that arise from ovarian surface cells, as has been proposed previously Oil the basis of studies of protein markers. (C) 2009 Elsevier Inc. All rights reserved.
Notes
Bendoraite, Ausra Knouf, Emily C. Garg, Kavita S. Parkin, Rachael K. Kroh, Evan M. O'Briant, Kathy C. Ventura, Aviva P. Godwin, Andrew K. Karlan, Beth Y. Drescher, Charles W. Urban, Nicole Knudsen, Beatrice S. Tewari, Muneesh Ovarian Cancer SPORE at FCCC [P50 CA083638]; Pacific Ovarian Cancer Research Consortium Ovarian SPORE [P50 CA83636]; Mary Kay Ash Charitable Foundation ; American Cancer Society California Division - Early Detection Professorship [SIOP-06-258-01-CCE]; Fred Hutchinson Cancer Research Center ; Canary Foundation We thank Anders Lund for providing the ZEB2 3' UTR luciferase construct, Dr. Guangwei Du for the pSM30 vector, Carolyn Slater for primary tissue culturing, Robin G. Tharakan for cell photography, Chris Tachibana for assistance in scientific writing, and Travis E. Mitchell for scientific administrative support. We thank Dr. N. Ueno for the Hey cell line. The study was supported in part by the Ovarian Cancer SPORE at FCCC (P50 CA083638), the Pacific Ovarian Cancer Research Consortium Ovarian SPORE (P50 CA83636 to K.C.O'B., A.P.V., B.Y.K, Ch.W. D., N.U., B.S.K., and M.T.), a Mary Kay Ash Charitable Foundation grant (to M.T.), the American Cancer Society California Division - Early Detection Professorship grant (SIOP-06-258-01-CCE to B.Y.K.), and by Fred Hutchinson Cancer Research Center and Canary Foundation funding (New Development funds to M.T.). 47 Academic press inc elsevier science; 525 b st, ste 1900, san diego, ca 92101-4495 usa 537jh