Thrombin promotes epithelial ovarian cancer cell invasion by inducing epithelial-mesenchymal transition.
10.3802/jgo.2013.24.3.265
- Author:
Yi Cun ZHONG
1
;
Ting ZHANG
;
Wen DI
;
Wei Ping LI
Author Information
1. Department of Obstetrics and Gynecology, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China. weiping_li61@yahoo.cn
- Publication Type:Original Article
- Keywords:
Epithelial-mesenchymal transition;
Epithelial ovarian cancer;
Invasion;
Thrombin
- MeSH:
Blotting, Western;
Cadherins;
Cytokines;
Down-Regulation;
Enzyme-Linked Immunosorbent Assay;
Epithelial-Mesenchymal Transition;
Hirudins;
Interleukin-6;
Interleukins;
Matrix Metalloproteinases;
Neoplasms, Glandular and Epithelial;
Ovarian Neoplasms;
Phenotype;
Polymerase Chain Reaction;
Prognosis;
Proteins;
RNA, Messenger;
Thrombin;
Up-Regulation;
Vimentin
- From:Journal of Gynecologic Oncology
2013;24(3):265-272
- CountryRepublic of Korea
- Language:English
-
Abstract:
OBJECTIVE: Over-expression of thrombin in ovarian cancer cells is associated with poor prognosis. In this study, we investigated the role of thrombin in inducing epithelial-mesenchymal transition (EMT) in SKOV3 epithelial ovarian cancer cells. METHODS: After thrombin treatment SKOV3 cells were subjected to western blots, reverse-transcription PCR, and enzyme-linked immunosorbent assay to quantify EMT-related proteins, mRNA expression of SMAD2, DKK1, and sFRP1, and the secretion of matrix metalloproteinases (MMPs) and cytokines. Meanwhile, invasion ability was evaluated using transwell assays. RESULTS: The results indicated a dose- and time-dependent down-regulation of E-cadherin and upregulation of N-cadherin and vimentin in thrombin-treated SKOV3 cells, compared with the thrombin-free control group (p<0.05). There was a dose- and time-dependent increase in the levels of SMAD2 and DKK1 mRNAs and a decrease in the levels of sFRP1 mRNA in thrombin-treated SKOV3 cells compared to control cells (p<0.05). Thrombin-treated SKOV3 cells exhibited increased secretion of MMP-9, MMP-2, interleukin (IL)-8, and IL-6 and increased invasion compared to untreated cells (p<0.05). Thrombin altered the morphology of SKOV3 cells to a spindle-like phenotype. Addition of hirudin to thrombin-treated cells reversed the effects of thrombin. CONCLUSION: Thrombin induced EMT and promoted the invasion of SKOV3 cells, possibly via distinct signaling pathways. Hirudin inhibited the effects of thrombin, suggesting that anticoagulant therapy could be a novel therapeutic strategy for ovarian carcinoma.
