Brief Reports
Copyright ©The Author(s) 2001. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Oct 15, 2001; 7(5): 726-731
Published online Oct 15, 2001. doi: 10.3748/wjg.v7.i5.726
Identification of differentially expressed genes in normal mucosa, adenoma and adenocarcinoma of colon by SSH
Min-Jie Luo, Mao-De Lai
Min-Jie Luo, Mao-De Lai Department of Pathology, School of Medicine, Zhejiang University, Hangzhou 310031, Zhejiang Province, China
Author contributions: All authors contributed equally to the work.
Supported by Science Foundation of the Education Department of Zhejiang Province
Correspondence to: Mao-De Lai, M.D., Department of Pathology, School of Medicine, Zhejiang University, Hangzhou 310031, Zhejiang Province, China lmd@sun.zju.edu.cn
Telephone: +86-571-87217134, Fax: +86-571-87951358
Received: July 5, 2001
Revised: August 6, 2001
Accepted: August 16, 2001
Published online: October 15, 2001
Abstract

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 × 102 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.

Keywords: colonic neoplasms/genetics, adenoma/genetics, adenocarcinoma/genetics, intestinal mucosa/metabolism, nucleic acid hybridization