Exome Sequencing of Normal and Isogenic Transformed Human Colonic Epithelial Cells (HCECs) Reveals Novel Genes Potentially Involved in the Early Stages of Colorectal Tumorigenesis.
Exome Sequencing of Normal and Isogenic Transformed Human Colonic Epithelial Cells (HCECs) Reveals Novel Genes Potentially Involved in the Early Stages of Colorectal Tumorigenesis.
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DOI：
10.1186/1471-2164-16-s1-s8
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发表时间：
2015
期刊：
影响因子：
4.4
通讯作者：
Shay JW
中科院分区：
文献类型：
--
作者：
Zhang L;Kim S;Jia G;Buhmeida A;Dallol A;Wright WE;Fornace AJ;Al-Qahtani M;Shay JW
我们已经产生了一系列同基因衍生的永生化人结肠上皮细胞（HCEC 1CT 和 HCEC 2CT）系，包括亲本非永生化正常细胞株。 CDK4和hTERT永生化结肠上皮细胞系（HCEC 1CT）最初是核型正常的二倍体，表达一系列上皮细胞标志物，包括干细胞标志物。在应激组织培养条件下，HCEC 1CT 系中发生自发的非整倍体事件，导致单个染色体变化，从而形成稳定的三体性 7 细胞系 (1CT7)。 7 三体性发生在约 40% 的人类良性腺瘤（息肉）中，因此二倍体 HCEC 1CT 细胞中的这种特定染色体变化似乎是非随机的。此外，我们通过引入野生型 APC 和 TP53 的稳定敲低，以及异位引入突变型 Krasv12 和突变型 APC (A1309)，对 HCEC 1CT 系进行了部分转化，这些都是结直肠癌 (CRC) 中常见的突变。进行全外显子组测序和生物信息学分析，以全面检查这些同基因细胞系的遗传背景。这些实验进展的细胞系的外显子组测序概括了先前报道的参与结直肠癌肿瘤发生的一系列基因。此外，测序揭示了在 1CT7 和 A1309 细胞中特异性检测到的一系列新基因，但在正常二倍体 1CT 细胞中未检测到。本研究证明了使用同基因实验衍生的 HCEC 系作为模型来概括 CRC 的发生和进展的效用。外显子组测序揭示了一系列可能在结直肠癌肿瘤发生中发挥重要作用的新基因。
We 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). Whole exome sequencing and bioinformatic analyses were performed to comprehensively examine the genetic background of these isogenic cell lines. Exome 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. This 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.