摘 要 目的：探讨Raf激酶抑制蛋白（Raf kinase inhibitor protein，RKIP）对卵巢癌SKOV-3细胞化疗敏感性的影响。方法：以脂质体法将含有人全长RKIP基因的真核表达质粒pcDNA3.1-ssRKIP转染入SKOV-3细胞中，Western blotting检测SKOV-3细胞中RKIP蛋白的表达。不同浓度顺铂作用转染后的SKOV-3细胞，MTS法观察RKIP基因转染对顺铂处理后SKOV-3细胞增殖的影响，流式细胞仪检测RKIP基因转染对顺铂诱导SKOV-3细胞凋亡及细胞周期的影响。结果：pcDNA3.1-ssRKIP转染的SKOV-3细胞RKIP表达明显升高。不同浓度顺铂处理细胞24、48、72 h后，RKIP基因转染细胞增殖抑制率显著高于对照细胞（P<0.05）。用2.5 μg/ml顺铂作用SKOV-3细胞24 h后，RKIP转染细胞的凋亡率为（10.86±0.73）%，明显高于未转染细胞的（4.27±0.67）%和空质粒转染细胞的（4.02±0.43）%（P<0.01)；在无顺铂作用情况下，RKIP转染细胞的凋亡率为（3.11±0.78）%，仍然高于未转染细胞的（1.51±0.13）%和转染空质粒细胞的（1.97±0.46）%（P<0.01)。细胞周期检测结果显示，RKIP转染细胞G0/G1期的比例下降，S期的比例增加，转染的SKOV-3细胞发生S期阻滞。结论：RKIP基因的转染可以增加卵巢癌SKOV-3细胞对化疗药物顺铂的敏感性。

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 G0/G1 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.

国家自然科学基金资助项目（No.30670801）；天津市科委基金资助项目（No.06YFJMJC08300）