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目的 用基因芯片技术研究高低转移人卵巢癌细胞系 (HO 8910PM和HO 8910 )基因表达谱差异 ,筛选与转移相关的基因。方法 按一步法分别抽提高低转移人卵巢癌细胞和对照正常卵巢组织的总RNA并纯化mRNA ;分别将等量的高低转移人卵巢癌细胞及对照组织的mRNA逆转录合成以Cy5和Cy3标记的cDNA一链做探针 ,分别混合后在 2张含有 4 0 96条双点人类全长基因的芯片上进行杂交。经洗片后用ScanArray 30 0 0扫描仪扫描芯片荧光信号图像 ,用ImaGene 3 0软件对扫描图象进行数字化处理 ,计算机分析高低转移人卵巢癌细胞系之间及与正常卵巢上皮基因表达谱差异。结果 人卵巢癌细胞系HO 8910细胞与正常卵巢上皮比较差异 3倍以上共有 35 5个基因。高转移人卵巢癌细胞系HO 8910PM细胞与正常卵巢上皮比较差异 3倍以上共有 32 3个基因。高转移人卵巢癌细胞系HO 8910PM与母系HO 8910比较差异 2倍以上共有 16 5个基因 ,其中两株细胞系比较差异 3倍以上共有 2 1个基因。结论 两株人卵巢癌细胞系与正常卵巢上皮细胞基因表达谱存在差异 ,提示这些基因与卵巢癌的发生和发展及高转移特性有关。本实验说明利用基因芯片对基因表达谱的检测可以为人体卵巢癌的基因诊断、治疗和预防提供新的方向。
OBJECTIVE: To study the gene expression profiling of human ovarian cancer cell lines (HO 8910PM and HO 8910) using gene chip technique to screen for genes involved in metastasis. Methods The total RNA was extracted from human ovarian cancer cells and control normal ovary tissues by one-step method, and the mRNA was purified. The mRNAs of ovarian cancer cells and control tissues with the same level were reverse transcribed to Cy5 and Cy3 The cDNA strand was used as a probe and hybridized separately on two chips containing 4096 double-stranded human full-length genes. After the chip was washed, the fluorescence signal of the chip was scanned by ScanArray 30 0 0 scanner. The scanned image was digitized with ImaGene 30 software. The gene expression profile of human ovarian cancer cell lines with and without metastasis was analyzed by computer. . Results The human ovarian cancer cell line HO-8910 cells had 355 genes more than 3-fold difference from normal ovarian epithelial cells. The highly metastatic human ovarian cancer cell line HO-8910PM had more than 3-fold differences from normal ovarian epithelial cells in a total of 32 3 genes. The highly metastatic human ovarian cancer cell line HO 8910PM had more than twice as many as the maternal HO 8910 with a total of 165 genes, of which two of the two cell lines had a difference of more than 3 fold, and a total of 21 genes. Conclusion The gene expression profiles of two human ovarian cancer cell lines and normal ovarian epithelial cells are different, suggesting that these genes are associated with the occurrence, development and metastasis of ovarian cancer. This experiment shows that the use of gene chip gene expression profiling can provide a new direction for gene diagnosis, treatment and prevention of human ovarian cancer.