Journal Article

Gene Expression Profiling of Cultured Human Bronchial Epithelial and Lung Carcinoma Cells

Gary M. Hellmann, Wanda R. Fields and David J. Doolittle

in Toxicological Sciences

Volume 61, issue 1, pages 154-163
Published in print May 2001 | ISSN: 1096-6080
Published online May 2001 | e-ISSN: 1096-0929 | DOI: http://dx.doi.org/10.1093/toxsci/61.1.154
Gene Expression Profiling of Cultured Human Bronchial Epithelial and Lung Carcinoma Cells

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Lung cancer is a complex collection of diseases that is thought to begin with single mutated progenitor cells and culminates in any of several clinically described pathologies. Our knowledge of the molecular events that lead to different lung cancer types—small cell carcinoma, squamous cell carcinoma, adenocarcinoma, and large cell carcinoma—is incomplete. Nonetheless, it is evident that genetic changes that impact multiple molecular networks are involved in the generation of each specific phenotype. Due to the obvious complexity of these processes, the simultaneous quantitative monitoring of changes in the expression of genes that define these networks can provide mechanistic information to increase our understanding of the molecular basis for human pulmonary carcinogenesis. To this end, we have employed a commercially available human cDNA array (AtlasTM Human Array, Clontech Laboratories) to systematically screen for alterations in the expression of 600 genes in normal human bronchial epithelial (NHBE) cells as well as in several lung carcinoma lines. Studies on the reproducibility and variability of array results indicate that a 2-fold or greater difference in the expression of a particular gene could be considered a real difference in transcript abundance. Accuracy of gene expression as measured in the array was verified by comparing mRNA levels of the proto-oncogene c-myc in the array with results obtained by traditional Northern blot analysis and by quantitative RT-PCR. Gene expression profiles were compared within and among cell types. The differential expression of 17 genes, including downregulation of MRP8 and MRP14 and upregulation of CYP1B1, was observed in all four carcinoma lines compared to NHBE cells. The direction of all 17 gene expression differences, either upregulation or downregulation relative to NHBE cells, was the same for all four carcinoma lines, underscoring their common molecular features. Each lung tumor line also exhibited a number of unique differences compared to both normal cells and the other tumor cell lines. These differences may be due to differences in the cellular origin and/or pathology of the cell lines studied.

Keywords: gene expression; array; lung carcinogenesis; mRNA; expression profiling; human cell culture; bronchial epithelial cells.

Journal Article.  6937 words.  Illustrated.

Subjects: Medical Toxicology ; Toxicology (Non-medical)

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