体外培养的不同亚型上皮性卵巢癌细胞系差异蛋白的分析

THE PRELIMINARY ANALYSIS OF DISTINCT PROTEINS FOR DIFFERENT SUBTYPES OF EPITHELIAL OVARIAN CANCER CELL LINES IN VITRO CULTURE

目的分析体外培养的不同亚型上皮性卵巢癌细胞系蛋白质表达差异,筛选卵巢癌的肿瘤标志物。方法提取体外培养的不同亚型上皮性卵巢癌细胞系(OVCAR-3、SKOV-3、3AO和TOV-112D)与原代培养的正常卵巢生发上皮细胞(OGE)的总蛋白,采用表面增强激光解吸离子化(SELDI)蛋白质芯片技术的H4和SAX2两种蛋白芯片检测其蛋白质谱,最后利用蛋白芯片阅读机(PBS-Ⅱ型)读取数据,获得的数据采用Ciphergen Proteinchip 3.0.2版本的软件分析。结果4种亚型的上皮性卵巢癌细胞系与正常卵巢生发上皮细胞比较,有16个蛋白发生明显改变,其中7个蛋白表达降低,9个蛋白表达升高。此外,不同亚型的上皮性卵巢癌细胞之间也有很多差异蛋白,尤其是OVCAR-3、SKOV-3和3AO三者与TOV-112D之间有18个蛋白仅在前3种癌细胞中表达,16个蛋白仅在后者表达。结论不同亚型上皮性卵巢癌细胞与正常卵巢生发上皮细胞的蛋白质谱相比较有共同的差异蛋白,同时不同亚型的上皮性卵巢癌细胞之间的蛋白质谱表达也有差别。

Objective To discover the common biomarkers in ovarian cancer cell lines and detect the distinct proteins for different types of ovarian cancer cell lines. Methods We applied SELDI proteinchip technique to detect total proteins of cell which were respectively derived from four kinds of epithelial ovarian cancer cell lines （OVCAR-3, SKOV-3, 3AO and TOV-112D） and ovarian germinal epithelium （OGE） in primary culture. Two different protein chips arrays, H4 and SAX2, were used in the process. H4 is a protein chip which captures low molecular weight peptides and proteins by hydrophobic interaction, and SAX2 captures proteins whose surface carries negative charges. Then, protein chips were examined in PBS-Ⅱ protein chip reader （Ciphergen Biosystem Inc） and the protein profiling was analyzed by protein chip Software 3.0.2. Results There were significant differences in sixteen peaks between the four epithelial ovarian cancer cell lines and OGE, seven （1236D, 1849D, 1871D, 2467D and 6652D captured by H4, 5280D and 9744D by SAX2） of which were down-regulated in epithelial ovarian cancer cells, and the others of which （5478D, 5646D, 6922D, 7038D, 7356D, 8291D, 9126D, 9334D and 10188D captured by H4） were up-regulated. Besides, there were many distinct proteins in the different subtypes of epithelial ovarian cancer cell, especially, eighteen of which were only detected in OVCAR-3, SKOV-3 and 3AO, and sixteen only in TOV-112D. Conclusion The results suggest that there are same distinct proteins in protein profiling between different ovarian cancer cells and normal ovarian germinal epithelium, and the discriminated proteins between different types of ovarian cancer cells are also found.