ALDH isozymes downregulation affects cell growth, cell motility and gene expression in lung cancer cellsReport as inadecuate




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Molecular Cancer

, 7:87

First Online: 24 November 2008Received: 19 June 2008Accepted: 24 November 2008

Abstract

BackgroundAldehyde dehydrogenase isozymes ALDH1A1 and ALDH3A1 are highly expressed in non small cell lung cancer. Neither the mechanisms nor the biologic significance for such over expression have been studied.

MethodsWe have employed oligonucleotide microarrays to analyze changes in gene profiles in A549 lung cancer cell line in which ALDH activity was reduced by up to 95% using lentiviral mediated expression of siRNA against both isozymes Lenti 1+3. Stringent analysis methods were used to identify gene expression patterns that are specific to the knock down of ALDH activity and significantly different in comparison to wild type A549 cells WT or cells similarly transduced with green fluorescent protein GFP siRNA.

ResultsWe confirmed significant and specific down regulation of ALDH1A1 and ALDH3A1 in Lenti 1+3 cells and in comparison to 12 other ALDH genes detected. The results of the microarray analysis were validated by real time RT-PCR on RNA obtained from Lenti 1+3 or WT cells treated with ALDH activity inhibitors. Detailed functional analysis was performed on 101 genes that were significantly different P < 0.001 and their expression changed by ≥ 2 folds in the Lenti 1+3 group versus the control groups. There were 75 down regulated and 26 up regulated genes. Protein binding, organ development, signal transduction, transcription, lipid metabolism, and cell migration and adhesion were among the most affected pathways.

ConclusionThese molecular effects of the ALDH knock-down are associated with in vitro functional changes in the proliferation and motility of these cells and demonstrate the significance of ALDH enzymes in cell homeostasis with a potentially significant impact on the treatment of lung cancer.

List of abbreviationsALDHaldehdye dehydrogenase

ALDH1A1 and ALDH3A1aldehyde dehydrogenase class 1A1 and 3A1

DEABdiethylaminobezaldehyde

TTtetraethylthiuram disulfide or disulfiram

NSCLCnon small cell lung cancer

Lenti 1 or Lenti 3cells expressing siRNA against ALDH1A1 or ALDH3A1 using lentivirus

Lenti 1+3cells expressing siRNA against ALDH1A1 and ALDH3A1

WTrefers to wild type cells

GFPrefers to cells expressing siRNA against the green fluorescence protein.

Electronic supplementary materialThe online version of this article doi:10.1186-1476-4598-7-87 contains supplementary material, which is available to authorized users.

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Author: Jan S Moreb - Henry V Baker - Lung-Ji Chang - Maria Amaya - M Cecilia Lopez - Blanca Ostmark - Wayne Chou

Source: https://link.springer.com/







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