title: TESTIN was commonly hypermethylated and involved in the epithelial-mesenchymal transition of endometrial cancer.
abstract: We previously reported frequent loss of TESTIN in human endometrial carcinoma, which significantly suppressed tumor proliferation and invasion. Herein, we further explored the mechanisms underlying TESTIN loss and its roles in the epithelial-mesenchymal transition (EMT, a key step for tumor spreading). Methylation-specific PCR was performed to investigate the promoter status of TESTIN in a panel of endometrial cancer and normal endometrium tissues. The expression of TESTIN mRNA was determined by real-time PCR. Up- and down-regulation of TESTIN were achieved by transient transfection with pcDNA3.1-TESTIN and shRNA-TESTIN plasmids, respectively. The EMT alterations were observed under the optical microscope and EMT-related markers were detected by real-time PCR and western blot. Compared to the control (3.6%), TESTIN was hypermethylated in 43.7% endometrial cancer tissues (p < 0.001). Moreover, TESTIN hypermethylation was significantly correlated with advanced tumor stage, deep myometrial invasion and lymphatic node metastasis. In vitro, the demethylating agent dramatically restored the expression of TESTIN. In addition, up-regulation of TESTIN significantly suppressed the EMT procedure; whereas down-regulation of TESTIN enhanced EMT. In conclusion, we demonstrated that loss of TESTIN was mainly caused by hypermethylation, which might be a potent prognostic marker. Furthermore, we proved that TESTIN significantly suppressed the EMT procedure, proposing restoration of TESTIN to be a novel therapeutic strategy for endometrial carcinoma.
authors.affiliation: Department of Reproductive Center, First Affiliated Hospital of Soochow University, Suzhou, Jiangsu Province, China; Department of Obstetrics and Gynecology, The Affiliated Hospital of Jiangnan University and the Fourth People's Hospital of Wuxi, Wuxi, Jiangsu Province, China.
authors.forename: Ruofan
authors.initials: R
authors.lastname: Dong
authors.forename: Hong
authors.initials: H
authors.lastname: Pu
authors.forename: Yuan
authors.initials: Y
authors.lastname: Wang
authors.forename: Jinjin
authors.initials: J
authors.lastname: Yu
authors.forename: Kuixian
authors.initials: K
authors.lastname: Lian
authors.forename: Caiping
authors.initials: C
authors.lastname: Mao
mesh_terms: D045744:Cell Line, Tumor; D003598:Cytoskeletal Proteins / Q000235:genetics*; D019175:DNA Methylation; D004273:DNA, Neoplasm / Q000737:chemistry / Q000235:genetics; D015536:Down-Regulation; D016889:Endometrial Neoplasms / Q000235:genetics* / Q000473:pathology*; D058750:Epithelial-Mesenchymal Transition / Q000235:genetics*; D005260:Female; D006801:Humans; D060588:LIM Domain Proteins / Q000235:genetics*; D008875:Middle Aged; D011379:Prognosis; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics; D016601:RNA-Binding Proteins; D014407:Tumor Cells, Cultured; D015854:Up-Regulation
chemical_list: D003598:Cytoskeletal Proteins; D004273:DNA, Neoplasm; D060588:LIM Domain Proteins; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; C429688:TES protein, human
keywords: EMT; Endometrial carcinoma; TESTIN; hypermethylation