Exome Sequencing of Normal and Isogenic Transformed Human Colonic Epithelial Cells (HCECs) Reveals Novel Genes Potentially Involved in the Early Stages of Colorectal Tumorigenesis

BMC Genomics

Research

Mohammed Al-Qahtani¹  Ashraf Dallol²  Albert J Fornace³  Lu Zhang⁴  Gaoxiang Jia⁴  Jerry W Shay⁴  Sang Bum Kim⁴  Abdelbaset Buhmeida⁴  Woodring E Wright⁴ 

[1] Center of Excellence in Genomic Medicine Research (CEGMR), King Fahad Medical Research Center, King Abdulaziz University, Jeddah, Kingdom of Saudi Arabia;Center of Excellence in Genomic Medicine Research (CEGMR), King Fahad Medical Research Center, King Abdulaziz University, Jeddah, Kingdom of Saudi Arabia;KACST Technology Innovation Center in Personalized Medicine, King Abdulaziz University, Jeddah, Kingdom of Saudi Arabia;Department of Biochemistiry and Molecular & Cellular Biology and Lombardi Comprehensive Cancer Center, Georgetown University, 20057, Washington, DC, USA;Department of Cell Biology, University of Texas Southwestern Medical Center, 5323 Harry Hines Boulevard, 75390-9039, Dallas, TX, USA;

关键词: A1309 Cell;    Exome Sequencing;    Colorectal Adenoma;    Exome Capture;    Human Colonic Epithelial Cell;   

DOI  :  10.1186/1471-2164-16-S1-S8

来源: Springer

【 摘 要 】

BackgroundWe have generated a series of isogenically derived immortalized human colonic epithelial cell (HCEC 1CT and HCEC 2CT) lines, including parental un-immortalized normal cell strains. The CDK4 and hTERT immortalized colonic epithelial cell line (HCEC 1CT) is initially karyotypically normal diploid and expresses a series of epithelial cell markers including stem cell markers. Under stressful tissue culture conditions, a spontaneous aneuploidy event occurred in the HCEC 1CT line, resulting in a single chromosomal change leading to a stable trisomy 7 cell line (1CT7). Trisomy 7 occurs in about 40% of all benign human adenomas (polyps) and thus this specific chromosomal change in diploid HCEC 1CT cells appears to be non random. In addition, we have partially transformed the HCEC 1CT line by introducing stable knockdown of wild type APC and TP53, and ectopically introducing a mutant Krasv12 and a mutant version of APC (A1309), all commonly found mutations in colorectal cancer (CRC).MethodsWhole exome sequencing and bioinformatic analyses were performed to comprehensively examine the genetic background of these isogenic cell lines.ResultsExome sequencing of these experimentally progressed cell lines recapitulates a list of genes previously reported to be involved in CRC tumorigenesis. In addition, sequencing revealed a collection of novel genes specifically detected in 1CT7 and A1309 cells but not normal diploid 1CT cells.ConclusionThis study demonstrates the utility of using isogenic experimentally derived HCEC lines as a model to recapitulate CRC initiation and progression. Exome sequencing reveals a collection of novel genes that may play important roles in CRC tumorigenesis.

【 授权许可】

Unknown   
© Zhang et al; licensee BioMed Central Ltd. 2015. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.