Colorectal Cancer
Copyright ©The Author(s) 2004. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. May 1, 2004; 10(9): 1301-1305
Published online May 1, 2004. doi: 10.3748/wjg.v10.i9.1301
Construction of a metastasis-associated gene subtracted cDNA library of human colorectal carcinoma by suppression subtraction hybridization
Li Liang, Yan-Qing Ding, Xin Li, Guang-Zhi Yang, Jun Xiao, Li-Chun Lu, Jin-Hua Zhang
Li Liang, Yan-Qing Ding, Xin Li, Guang-Zhi Yang, Li-Chun Lu, Jin-Hua Zhang, Department of Pathology, the First Military Medical University, Guangzhou 510515, Guangdong Province, China
Jun Xiao, Department of Tropic Medicine, the First Military Medical University, Guangzhou 510515, Guangdong Province, China
Author contributions: All authors contributed equally to the work.
Supported by Military Medical Foundation of China, No. 01MA128
Correspondence to: Yan-Qing Ding, Professor, Department of Pathology, the First Military Medical University, Guangzhou 510515, Guangdong Province, China. wcjd@public.bta.net.cn
Telephone: +86-20-61642148 Fax: +86-20-61642148
Received: August 26, 2002
Revised: March 20, 2003
Accepted: April 11, 2003
Published online: May 1, 2004
Abstract

AIM: To construct a differentially-expressed gene subtracted cDNA library from two colorectal carcinoma (CRC) cell lines with different metastatic phenotypes by suppression subtractive hybridization.

METHODS: Two cell lines of human CRC from the same patient were used. SW620 cell line showing highly metastatic potential was regarded as tester in the forward subtractive hybridization, while SW480 cell line with lowly metastatic potential was treated as tester in the reverse hybridization. Suppression subtractive hybridization (SSH) was employed to obtain cDNA fragments of differentially expressed genes for the metastasis of CRC. These fragments were ligated with T vectors, screened through the blue-white screening system to establish cDNA library.

RESULTS: After the blue-white screening, 235 white clones were picked out from the positive-going hybridization and 232 from the reverse. PCR results showed that 200-700 bp inserts were seen in 98% and 91% clones from the forward and reverse hybridizations, respectively.

CONCLUSIONS: A subtractive cDNA library of differentially expressed genes specific for metastasis of CRC can be constructed with SSH and T/A cloning techniques.

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