【摘要】 目的顺铂对细胞DNA链作用具有选择性,它对增生活跃的细胞,特别是S期细胞杀伤力强。将雌激素先作用卵巢癌细胞一定时间后再加入顺铂,判断它能否作为促使癌细胞进入顺铂敏感期诱导剂并初步探讨雌激素协同顺铂的作用机制。方法选择人卵巢腺癌上皮细胞系COC1和顺铂耐药系COC1/DDP为研究对象。MTT法先测定预先加入雌激素16h对10μmol/L顺铂作用下的卵巢癌细胞生长影响。然后利用双向凝胶电泳分离预先加入50μmol/L雌激素16h与10μmol/L顺铂共同培养COC1/DDP和COC1细胞48h的总蛋白,银染显色,Imagemaster图像分析系统分析,从胶中选取部份分离较好的差异蛋白质点,基质辅助激光解析电离飞行时间质谱分析获取肽质量指纹图谱,Peptident软件搜索数据库鉴定蛋白质。结果雌激素促进顺铂对卵巢癌细胞系COC1和COC1/DDP生长有抑制作用。获取了重复性和分辨率较好的50μmol/L雌激素与10μmol/L顺铂共同培养的COC1和COC1/DDP细胞双向凝胶电泳图谱。质谱分析了5个蛋白质点,获取了5张肽质量指纹图谱。4个可能参与雌激素促进顺铂效应蛋白质:COC1中有锌指蛋白多型2和FK506结合蛋白5;COC1/DDP中为微管β链蛋白和胞核包含颉酪肽蛋白相似蛋白。结论雌激素增加了卵巢癌细胞系COC1/DDP顺铂敏感性。雌激素协同顺铂抑制COC1和COC1/DDP细胞生长中出现差异蛋白质对理解雌激素提高卵巢癌顺铂敏感性机制提供了有价值的信息。更多还原

【Abstract】 Objective To increase the efficacy of chemotherapy of ovarian cancer by increasing the sensitivity of tumour cell to cisplatin. Cisplatin is known to selectively act on DNA of proliferative cells, especially the cells in S-phase. This study aims to examine if estrogen, when added before cisplatin treatment, can increase the sensitivity of ovarian cancer cell to cisplatin. The mechanism of interaction between estrogen and cisplatin was also study. Method Two human ovarian cancer cell lines, namely the cisplatin-sensitive line COC1 and the cisplatin-resistant line COC1/DDP, were used. Effects of estrogen pretreatment on the cell growth were determined by MTT reduction method. Two-dimensional polyacrylamide gel electrophoresis (2-DE) was used to separate the total cellular proteins of estrogen (50 μmol/L) and cisplatin (10 μmol/L) treated COC1 and COC1/DDP cells. Protein spots were stained by silver staining, and the profile was analyzed by ImageMaster 2-DE analysis software. Peptides was identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry(MALDI-TOF-MS). Result Estrogen pretreatment increased the growth inhibitory activity of cisplatin in both COC1 and COC1/DDP cell lines. High resolution 2-DE profiles form estrogn and cisplatin treated COC1 and COC1/DDP ovarian cancer cell lines were obtained. A total of five protein spots were analyzed by MALDI-TOF-MS, and 5 peptide mass fingerprints (PMF) were obtained. Four proteins were identified. These included ZFPM-2 (Zinc finger protein multitype 2) and FK506-BP5 (FK506-binding protein 5) from COC1 cell, and TB-β (Tubulin beta chain) and NVLp (Nuclear VCP-like protein) from COC1/DDP cell line. Conclusion Estrogen can increase the cisplatin-sensitivity in resistant COC1/DDP cell. Further studies on the proteins induced during inhibition of cell-growth induced by estrogen and cisplatin may enhance our understanding on the mechanism of cisplatin-sensitivity in ovarian cancer.更多还原