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Analysis of target genes induced by IL-13 cytotoxin in human glioblastoma cells

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Abstract

IL-13 cytotoxin comprised of IL-13 and a mutated form of Pseudomonas exotoxin (fusion protein termed IL-13-PE38QQR) has been shown to inhibit protein synthesis leading to necrotic and apoptotic cell death in glioblastoma cells that express high levels of interleukin-13 receptors (IL-13R). To identify target genes of cell death and other cellular genes with IL-13 receptors in glioblastoma cells, we utilized the cDNA microarrays to analyze global gene expression profiles after IL-13 cytotoxin and IL-13 treatment. IL-13 cytotoxin mediated cytotoxicity to U251 cells in a dose-dependant manner. Hierarchical cluster analysis of differentially expressed genes in U251 glioma cells at different time points after IL-13 cytotoxin treatment showed three major groups, each representing a specific expression pattern. Randomly selected differentially expressed genes from each group were confirmed by RT-PCR analysis. Most down-regulated genes belong to cell adhesion, motility, angiogenesis, DNA repair, and metabolic pathways. While up-regulated genes belong to cell cycle arrest, apoptosis, signaling and various metabolic pathways. Unexpectedly, at early time points, both IL-13 and IL-13 cytotoxin induced several genes belonging to different pathways most notably IL-8, DIO2, END1, and ALDH1A3 indicating that these genes are early response genes and their products may be associated with IL-13R. In addition, IL-13 cytotoxin induced IL-13Rα2 mRNA expression during the treatment in glioma cells. Our results indicate that novel cellular genes are involved with IL-13 receptors and that IL-13 cytotoxin induced cell death involves various target genes in human glioblastoma cells. On going studies will determine the role of associated genes and their products in the IL-13R functions in glioma cells.

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  1. Laboratory of Molecular Tumor Biology, CBER/NCI Genomics Program, Division of Cellular and Gene Therapies, Center for Biologics Evaluation and Research, Food and Drug Administration, CBER/FDA, NIH Bldg 29B, Rm 2NN10, 29 Lincoln Dr., Bethesda, MD, USA, 20892

    Jing Han & Raj K. Puri

  2. Bioinformatics & Molecular Analysis Section, Center for Information Technology, National Institutes of Health, Bethesda, MD, USA

    Liming Yang

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  1. Jing Han
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Han, J., Yang, L. & Puri, R.K. Analysis of target genes induced by IL-13 cytotoxin in human glioblastoma cells. J Neurooncol 72, 35–46 (2005). https://doi.org/10.1007/s11060-004-3119-7

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