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Raf kinase inhibitor protein enhances chemosensitivity of ovarian cancer cells
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Abstract:
Abstract Objective:To explore the effect of Raf kinase inhibitor protein (RKIP) on the chemosensitivity of ovarian cancer SKOV-3 cells. Methods: Eukaryotic expression plasmid pcDNA3.1-ssRKIP containing full-length human RKIP cDNA was transfected into ovarian cancer cell line SKOV-3 by lipofect assay. Expression of RKIP in SKOV-3 cells was determined by Western blotting analysis. pcDNA3.1-ssRKIP-transfected SKOV-3 cells were treated with different concentrations of cisplatin, and the effect of RKIP on the proliferation of SKOV-3 cells treated with cisplatin was measured by MTS assay. Flow cytometry was used to detect the effect of RKIP on changes of apoptosis and cell cycle of SKOV-3 cells after cisplatin treatment. Results: The expression of RKIP in SKOV-3 cells was significantly increased after transfection with pcDNA3.1-ssRKIP. The growth inhibitory rate of SKOV-3 cells in pcDNA3.1-ssRKIP transfection group was significantly higher than that in the control group after treatment with different concentrations of cisplatin for 24 h, 48h or 72 h (P<0.05). After treatment with cisplatin at 2.5 μg/ml for 24 hours, pcDNA3.1-ssRKIP-transfected SKOV-3 cells showed a significantly higher percentage of apoptosis (10.86±0.73)% than non-transfected cells (4.27±0.67)% and empty vector-transfected cells (4.02±0.43)%. Without cisplatin treatment, the percentage of apoptosis for SKOV-3 cells transfected with pcDNA3.1-ssRKIP was (3.11±0.78)%, which was significantly higher than those of the non-transfected cells (1.51±0.13)% and empty vector-transfected cells (1.97±0.46)%. After cisplatin treatment, there were fewer cells in G0/G1 phase and more cells in S phase in pcDNA3.1-ssRKIP-transfected cells compared with the control cells, suggesting that cisplatin caused more S phase arrest in transfected cells. Conclusions: Over-expression of RKIP gene can increase chemosensitivity of ovarian cancer SKOV-3 cells to cisplatin.
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Project Supported:
Project supported by the National Natural Science Foundation of China (No. 30670801), and the Foundation from Tianjin Municipal Science and Technology Commission （No. 06YFJMJC08300）
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