Raf激酶抑制蛋白增强卵巢癌细胞的化疗敏感性  

Raf kinase inhibitor protein enhances chemosensitivity of ovarian cancer cells

作　　者：宋继文[1] 高燕[2] 林璨璨[1] 李宏钊[1] 姚智[1] 邓为民[1] 

机构地区：[1]天津医科大学免疫学教研室,天津300070 [2]天津市中心妇产科医院妇产科,天津300070

出　　处：《中国肿瘤生物治疗杂志》2010年第2期155-160,共6页Chinese Journal of Cancer Biotherapy

基　　金：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)~~

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、72h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2.5μg/ml顺铂作用SKOV-3细胞24h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G0/G1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。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 eisplatin 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 eisplatin 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 transfeetion group was significantly higher than that in the control group after treatment with different concentrations of cisptatin 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-transfeeted cells （4.27± 0.67 ） % and empty vector-transfected cells （4.02 ± 0.43 ） %. Without eisplatin treatment, the percentage of apoptosis for SKOV-3 cells transfeeted 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-transfeeted cells （ 1.97 ± 0.46） %. After eisplatin 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.

关 键 词：RAF激酶抑制蛋白 卵巢癌 顺铂 化疗敏感性 

分 类 号：R735.7[医药卫生—肿瘤]