摘要
Objectives To study the difference between gene expressions of high (H0-8910PM) and low (HO-8910) metastatic human ovarian carcinoma cell lines and screen novel associated genes by cDNA microarray. Methods cDNA retro-transcribed from equal quantities of mRNA derived from high and low metastatic tumor cells or normal ovarian tissues were labeled with Cy5 and Cy3 fluorescein as probes. The mixed probe was hybridized with two pieces of BioDoor 4096 double dot human whole gene chip and scanned with a ScanArray 3000 laser scanner. The acquired image was analyzed by ImaGene 3.0 software. Results A total of 355 genes with expression levels more than 3 times larger were found by comparing the HO-8910 cell with normal ovarian epithelial cells. A total of 323 genes with expression levels more than 3 times larger in HO-8910PM cells compared to normal ovarian epithelium cells were also detected. A total of 165 genes whose expression levels were more than two times those of HO-8910PM cells compared to their mother cell line (HO-8910) were detected. Twenty-one genes with expression levels 】3 times were found from comparison of these two tumor cell lines.Conclusions cDNA microarray techniques are effective in screening differential gene expression between two human ovarian cancer cell lines (H0-8910PM; HO-8910) and normal ovarian epithelial cells. These genes may be related to the genesis and development of ovarian carcinoma. Analysis of the human ovarian cancer gene expression profile with cDNA microarray may help in gene diagnosis, treatment and prevention.
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。
