人直肠腺癌消减杂交差异片段的生物信息学分析

Bioinformatics analysis of differential fragment of suppression subtractive hybridization in human rectum adenocarcinoma

目的探测人直肠腺癌发生和发展过程中差异表达的基因片段,了解人直肠腺癌的分子生物学机制。方法应用抑制性消减杂交技术,获得人直肠腺癌差异表达的基因,用生物信息学cDNA文库筛选的方法,对人直肠腺癌cDNA消减文库中的差异基因片段的序列作预测和分析。结果用抑制性消减杂交技术随机挑取获得的阳性克隆,提取质粒,双酶切分析,进行序列测定,得到与直肠腺癌发病相关的差异基因片段-9cDNA(Genbank登录号为BM360862),用cDNA文库筛选得到一个全长cDNA序列。基因表达系列分析(SAGE)显示除了直肠腺癌外、9cDNA还分布于前列腺腺癌、脑干成神经管细胞瘤、胰腺腺癌、胚胎正常血管内皮细胞、脑室管细胞瘤。结论直肠腺癌的发病是多环节和多步骤的过程,9cDNA与直肠腺癌发病相关,对9cDNA基因差异片段进一步研究,将为了解直肠腺癌发病的特异相关基因奠定基础。

Aim To detect the gene fragment of differential expression during the development and progression of human rectum adenocarcinoma so as to clarify the molecular mechanisms of human rectum adenocarcinoma. Methods Suppression subtractive hybridization (SSH) was used to obtain the ESTs sequence from the normal human rectum and human rectum adenocarcinoma (HRAC).The differential expression of ESTs was then predicted and analyzed by means of cDNA library screening of bioinformatics.Results The differential fragment of 9cDNA(Gene Bank number BM360862) was obtained by suppression subtractive hybridization, which selected positive clone randomly, extracted plasmid, sequenced and analyzed homology.The whole cDNA of differential fragment was obtained by means of cDNA library screening of bioinformatics.SAGE showed that 9cDNA fragment was also expressed in the carcinoma of prostate, the medullobastoma of brain stem, the pancreas adenocarcinoma, the embryo endothelial cell of blood and the neoplasm of cerebral ventricle.Conclusion The mechanisms of human rectum adenocarcinoma include multiplied links and steps and the 9cDNA fragment is related to human rectum adenocarcinoma.The further studies focusing on the 9cDNA fragment make the foundation for screening and cloning new specific oncogenes or tumor suppressor genes in human rectum adenocarcinoma.