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World J Gastroenterol. Sep 14, 2007; 13(34): 4620-4625
Published online Sep 14, 2007. doi: 10.3748/wjg.v13.i34.4620
Changes in gene-expression profiles of colon carcinoma cells induced by wild type K-ras2
Hong Li, Hou-Fa Cao, Yuan Li, Mei-Ling Zhu, Jun Wan
Hong Li, Hou-Fa Cao, Inpatient Department of Special Need Treatment, General Hospital of Chinese PLA, Beijing 100083, China
Yuan Li, Mei-Ling Zhu, Jun Wan, Department of South Building Gastroenterology, General Hospital of Chinese PLA, Beijing 100853, China
Author contributions: All authors contributed equally to the work.
Supported by National Natural Science Foundation of China, No. 30200326
Correspondence to: Jun Wan, Department of South Building Gastroenterology, General Hospital of Chinese PLA, Beijing 100083, China. wanjun@301hospital.com.cn
Telephone: +86-10-66937584 Fax: +86-10-66937584
Received: March 15, 2007
Revised: June 20, 2007
Accepted: June 23, 2007
Published online: September 14, 2007
Abstract

AIM: To further elucidate the possible molecular biological activity of wild type K-ras2 gene by detecting changes in wild type K-ras2 gene-induced gene-expression profiles of colon carcinoma cells using cDNA microarray techniques.

METHODS: Total RNA was isolated from peripheral blood of health volunteers. Reverse transcription of RNA and polymerase chain reaction were used to synthesize wild type K-ras2 cDNA. K-ras2 cDNA fragment was cloned into a T easy vector and sequenced. A eukaryotic expression vector pCI-neo-K-ras2 was constructed and transfected to Caco2 cell line using the liposome method. Finally, mRNA was isolated, reverse-transcribed to cDNA from pCI-neo-K-ras2 or pCI-neo blank vector-transfected Caco cells, and analyzed by cDNA microarray assay.

RESULTS: Restriction enzyme analysis and DNA sequencing verified that the constructed expression vector was accurate. High-quality RNA was extracted and reverse transcribed to cDNA for microarray assay. Among the 135 genes, the expression was up-regulated in 24 and down-regulated in 121. All these differentially expressed genes were related to cell proliferation, differentiation, apoptosis and signal transduction.

CONCLUSION: Differentially expressed genes can be successfully screened from wild type K-ras2-transfected colon carcinoma cells using microarray techniques. The results of our study suggest that wild type K-ras2 is related to the negative regulation of cell proliferation, metabolism and transcriptional control, and provide new clues to the further elucidation of its possible biological activity.

Keywords: Wild type K-ras2; Colon cancer; Microarray