基因芯片研究卵巢癌细胞系HO-8910差异基因表达

The research of the genes expression difference of ovarian carcinoma cell line(H0-8910)by cDNA microarray

目的 用基因芯片技术研究人卵巢癌细胞系HO -8910和正常卵巢上皮的基因表达谱 ,分析其基因表达变化 ,探讨卵巢癌发生、发展过程中与肿瘤相关的基因群及其功能 ,有助于从分子水平全面了解细胞癌变机理 ,为卵巢癌的临床诊断和治疗提供分子标记和靶基因。 方法 用一步法抽提人卵巢癌细胞和对照正常卵巢组织的总RNA并纯化mRNA ;将等量的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链做探针 ,混合后在含有4096条双点人类全长基因的芯片上进行杂交。经洗片后用ScanArray3000扫描仪扫描芯片荧光信号图像 ,用ImaGene3.0软件对扫描图像进行数字化处理 ,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355个基因 ,其中表达上调>3倍有88个 ,表达下调<0.33有267个。差异6倍以上共有75个基因 ,其中表达上调>6倍有13个 ,表达下调<0.17有62个。结论 通过基因谱平行比较表明 :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 :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355个基因。提示这些基因与卵巢癌的发生和发展有关。

Objective To study the genes expression of ovarian carcinoma cell line(H0-8910)and normal ovarian epithelium and identify as well as screen novel associated genes by cDNA microarray. Methods The cDNA retro-transcribed from equal quantity mRNA derived from tissues of ovarian carcinoma cell line or normal ovarian,which were labeled with Cy5and Cy3fluorescence as probes.The mixed prober were hybridizd with BioDoor4096double dot human whole gene chip.It was scanned by ScanArray3000laser scanner.The acquired image was analyzed by ImaGene3.0software. Results By applying the cDNA microarray we found:Total355genes whose expression level were larger than3times in HO-8910cell and normal ovarian epithelium cell were screened out,among these gene up>3and down regulated<0.33were88and267.In among these75genes whose exˉpression level were larger than6times in HO-8910cell and normal ovarian epitheliumcell were screened out,among these gene up>6and down regulated<0.17were13and62individual,and1newgene were found. Conclusion cDNA microarray technique is effective in screening the differentially exˉpressed genes between human ovarian cancer cell line(H0-8910)and normal ovarian epithelium cell.Total355genes whose expression level were larger than3times in HO-8910cell and normal ovarian epitheliumcell were screened out.It suggestion that these genes associated with occurrence,development and metastasis of ovarian carcinoma.Using the cDNA microarray to analysis of human ovarian cancer cell line gene expression profile difference will be helpful for the gene diagnosis,treatment and prevention and providing the newdirection.