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.
Subject(s)
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 , VimentinABSTRACT
Objective To study the relativity between serum vascular endothelial growth factor(sVEGF),its receptors(sVEGFR1 and sVEGFR2) and uterine myoma. Methods Thirty hospitalized patients with uterine myoma were chosen as the study group and the other healthy women of the same age as control group.The levels of sVEGF,sVEGFR1 and sVEGFR2 of the two groups were determined by using ELISA technology. Results The levels of sVEGF and sVEGFR2 in the study group were much higher than in the control group(P0.05).The levels of sVEGF and sVEGFR2 in women with myoma had no differences during the menstrual cycle.The level of sVEGFR2 was correlated with the number of myoma(P