Identification of differentially expressed genes in normal mucosa,adenoma and adenocarcinoma of colon by SSH
Identification of differentially expressed genes in normal mucosa,adenoma and adenocarcinoma of colon by SSH作者机构：Department of Pathology School of Medicine Zhejiang University Hangzhou 310031 China.
出 版 物：《World Journal of Gastroenterology》 (世界胃肠病学杂志（英文版）)
年 卷 期：2001年第7卷第5期
页 面：726-731页
核心收录：
学科分类：1002[医学-临床医学] 100214[医学-肿瘤学] 10[医学]
基 金：This study is supported by Science Foundation of the Education Department of Zhejiang Province
主 题：Gene Expression Regulation, Neoplastic Adenocarcinoma Adenoma Blotting, Northern Colorectal Neoplasms Gene Library Genetic Markers Humans Intestinal Mucosa Polymerase Chain Reaction RNA, Messenger Research Support, Non-U.S. Gov't
摘 要：AIM: To construct subtracted cDNA libraries and further identify differentially expressed genes that are related to the development of colorectal carcinoma(CRC). METHODS: Suppression subtractive hybridization(SSH) was done on cDNAs of normal mucosa, adenoma and adenocarcinoma tissues from the same patient. Three subtracted cDNA libraries were constructed and then hybridized with forward and backward subtracted probes for differential screening. Positive clones from each subtracted cDNA library were selected for sequencing and BLAST analysis. Finally, virtual Northern Blot confirmed such differential expression. RESULTS: By this way, there were about 3-4 X 10(2) clones identified in each subtracted cDNA library, in which about 85% positive clones were differentially screened. Sequencing and BLAST homology search revealed some clones containing sequences of known gene fragments and several possibly novel genes showing few or no sequence homologies with any known sequences in the database. CONCLUSION: All results confirmed the effectiveness and sensitivity of SSH. The differentially expressed genes during the development of CRC can be used to shed light on the pathogenesis of CRC and be useful genetic markers for early diagnosis and therapy.
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