title: The role of WWOX tumor suppressor gene in the regulation of EMT process via regulation of CDH1-ZEB1-VIM expression in endometrial cancer.
abstract: This study defines the role of WWOX in the regulation of epithelial to mesenchymal transition. A group of 164 endometrial adenocarcinoma patients was studied as well as an ECC1 well-differentiated steroid-responsive endometrial cell line, which was transducted with WWOX cDNA by a retroviral system. The relationship between WWOX gene and EMT marker (CDH1, VIM, ZEB1, SNAI1) expression on mRNA (RT-qPCR) and protein levels (western blotting) was evaluated. The EMT processes were also analysed in vitro by adhesion of cells to extracellular matrix proteins, migration through a basement membrane, anchorage-independent growth and MMP activity assay. DNA microarrays (HumanOneArray™) were used to determine WWOX-dependent pathways in an ECC1 cell line. A positive correlation was observed between WWOX and ZEB1, and a negative correlation between CDH1 and VIM. WWOX expression was found to inversely correlate with the risk of recurrence of tumors in patients. However, in the WWOX-expressing ECC1 cell line, WWOX expression was found to be inversely related with VIM and positively with CDH1. The ECC1/WWOX cell line variant demonstrated increased migratory capacity, with increased expression of metalloproteinases MMP2/MMP9. However, these cells were not able to form colonies in suspension and revealed decreased adhesion to fibronectin and fibrinogen. Microarray analysis demonstrated that WWOX has an impact on the variety of cellular pathways including the cadherin and integrin signalling pathways. Our results suggest that the WWOX gene plays a role in the regulation of EMT processes in endometrial cancer by controlling the expression of proteins associated with cell motility, thus influencing tissue remodeling, with the suppression of mesenchymal markers.
authors.affiliation: Department of Molecular Carcinogenesis, Medical University of Lodz, PL 90-752 Lodz, Poland.
authors.forename: Elżbieta
authors.initials: E
authors.lastname: Płuciennik
authors.affiliation: Department of Molecular Carcinogenesis, Medical University of Lodz, PL 90-752 Lodz, Poland.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Nowakowska
authors.affiliation: Department of Molecular Carcinogenesis, Medical University of Lodz, PL 90-752 Lodz, Poland.
authors.forename: Karolina
authors.initials: K
authors.lastname: Pospiech
authors.affiliation: Laboratory of Clinical and Transplant Immunology and Genetics, Copernicus Memorial Hospital in Lodz, PL 90-752 Lodz, Poland.
authors.forename: Anna
authors.initials: A
authors.lastname: Stępień
authors.affiliation: Bio-Tech Consulting Sp. z o.o, Faculty of Biomedical Science and Postgraduate Education, Medical University of Lodz, PL 90-752 Lodz, Poland.
authors.forename: Mateusz
authors.initials: M
authors.lastname: Wołkowicz
authors.affiliation: Faculty of Biomedical Sciences and Postgraduate Education, Medical University of Lodz, PL 90-752 Lodz, Poland.
authors.forename: Małgorzata
authors.initials: M
authors.lastname: Gałdyszyńska
authors.affiliation: Faculty of Biomedical Sciences and Postgraduate Education, Medical University of Lodz, PL 90-752 Lodz, Poland.
authors.forename: Marta
authors.initials: M
authors.lastname: Popęda
authors.affiliation: Clinical Division of Gynecological Oncology, Medical University of Lodz, PL 90-752 Lodz, Poland.
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Wójcik-Krowiranda
authors.affiliation: Clinical Division of Gynecological Oncology, Medical University of Lodz, PL 90-752 Lodz, Poland.
authors.forename: Andrzej
authors.initials: A
authors.lastname: Bieńkiewicz
authors.affiliation: Department of Molecular Carcinogenesis, Medical University of Lodz, PL 90-752 Lodz, Poland.
authors.forename: Andrzej K
authors.initials: AK
authors.lastname: Bednarek
mesh_terms: D015703:Antigens, CD; D015820:Cadherins / Q000235:genetics / Q000378:metabolism; D002448:Cell Adhesion; D045744:Cell Line, Tumor; D002465:Cell Movement; D016889:Endometrial Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D058750:Epithelial-Mesenchymal Transition; D005260:Female; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D010088:Oxidoreductases / Q000235:genetics* / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D025521:Tumor Suppressor Proteins / Q000235:genetics* / Q000378:metabolism*; D014746:Vimentin / Q000235:genetics / Q000378:metabolism; D000074584:WW Domain-Containing Oxidoreductase; D000071799:Zinc Finger E-box-Binding Homeobox 1
chemical_list: D015703:Antigens, CD; C513384:CDH1 protein, human; D015820:Cadherins; D018398:Homeodomain Proteins; D014157:Transcription Factors; D025521:Tumor Suppressor Proteins; D014746:Vimentin; C486307:ZEB1 protein, human; D000071799:Zinc Finger E-box-Binding Homeobox 1; D010088:Oxidoreductases; D000074584:WW Domain-Containing Oxidoreductase; C486071:WWOX protein, human