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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。
Objective To detect the gene fragments differentially expressed in the development and progression of human rectal adenocarcinoma and to understand the molecular biological mechanism of human rectal adenocarcinoma. Methods Suppression subtractive hybridization (SSH) technique was used to obtain differentially expressed genes in human rectal adenocarcinoma. The sequences of differentially expressed genes in cDNA subtractive library of human rectal adenocarcinoma were predicted and analyzed by bioinformatics cDNA library screening. Results The positive clones were randomly picked by suppression subtractive hybridization. Plasmids were extracted and double enzyme digestion analysis was carried out. Sequencing results showed that the difference gene fragment-9 cDNA (GenBank Accession No. BM360862) Library screening results in a full-length cDNA sequence. In addition to rectal adenocarcinoma, 9cDNA was also found in prostate adenocarcinoma, brainstem medulloblastoma, pancreatic adenocarcinoma, embryonic normal vascular endothelial cells, and ventricular tubular cell tumors. Conclusion The pathogenesis of rectal adenocarcinoma is a multi-step and multistep process. 9cDNA is associated with the pathogenesis of rectal adenocarcinoma. Further study on the 9cDNA gene fragments will lay the foundation for understanding the specific genes involved in the pathogenesis of rectal adenocarcinoma.