Gene expression profile of human colon cancer cells treated with cross-reacting material 197, a diphtheria toxin non-toxic mutant.

Abstract

Cross-Reacting Material 197 (CRM197) is a diphtheria toxin non-toxic mutant that has shown antitumor activity in mice and humans. It is still unclear whether this anti-tumorigenic effect depends on its strong inflammatory-immunological property, its ability to inhibit heparin-binding epidermal growth factor (HB-EGF), or even its possible weak toxicity. CRM197 is utilized as a specific inhibitor of HB-EGF that competes for the epidermal growth factor receptor (EGFR), overexpressed in colorectal cancer and implicated in its progression. In this study we evaluate the effects of CRM197 on HT-29 human colon cancer cell line behaviour and, for CRM197 recognized ability to inhibit HB-EGF, its possible influence on EGFR activation. In particular, while HT-29 does not show any reduction of viability after CRM197 treatment (MTT modified assay), or changes in cell cycle distribution (flow cytometry), in EGFR localization, phospho-EGFR detected signals (immunohistochemistry) or in morphology (scanning electron microscopy, SEM) they show a change in the gene expression profile by microarray analysis (cDNA microarray SS-H19k8). The overexpression of genes like protein phosphatase 2, catalytic subunit, alpha isozyme (PPP2CA), guanine nucleotide-binding protein G subunit alpha-1(GNAI1) and butyrophilin, subfamily 2, member A1 (BTN2A1) has been confirmed with real-time-qPCR. This is the first study where the CRM197 treatment on HT-29 shows a possible scarce implication of endogenous HB-EGF on EGFR expression and cancer cell development. At the same time, our results show the alteration of a specific and selected number of genes.

• Authors: S Rivetti, M Lauriola, M Voltattorni, M Bianchini, D Martini, C Ceccarelli, A Palmieri, G Mattei, M Franchi, G Ugolini, G Rosati, I Montroni, M Taffurelli, Rossella Solmi
• Journal: International journal of immunopathology and pharmacology (Int J Immunopathol Pharmacol) 2011 Jul-Sep Vol. 24 Issue 3 Pg. 639-49 ISSN: 0394-6320 [Print] England
• PMID: 21978696 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Bacterial Proteins
• Coloring Agents
• DNA, Complementary
• HBEGF protein, human
• Hbegf protein, mouse
• Heparin-binding EGF-like Growth Factor
• Intercellular Signaling Peptides and Proteins
• RNA, Neoplasm
• CRM197 (non-toxic variant of diphtheria toxin)
• ERBB4 protein, human
• ErbB Receptors
• Erbb4 protein, mouse
• Receptor, ErbB-4
• Trypan Blue

Topics

• Bacterial Proteins (pharmacology)
• Cell Cycle (drug effects)
• Cell Survival (drug effects)
• Colonic Neoplasms (genetics)
• Coloring Agents
• DNA, Complementary (biosynthesis, genetics)
• Enterocytes (drug effects, metabolism)
• ErbB Receptors (biosynthesis, genetics)
• Flow Cytometry
• Gene Expression Profiling (methods)
• Gene Expression Regulation, Neoplastic (drug effects)
• HT29 Cells
• Heparin-binding EGF-like Growth Factor
• Humans
• Immunohistochemistry
• In Situ Hybridization
• Intercellular Signaling Peptides and Proteins (metabolism)
• Microarray Analysis
• Microscopy, Electron, Scanning
• RNA, Neoplasm (biosynthesis, genetics)
• Receptor, ErbB-4
• Reverse Transcriptase Polymerase Chain Reaction
• Trypan Blue