title: 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: Dipartimento di Istologia, Embriologia e Biologia Applicata, Universit&#x00E0; di Bologna, Bologna, Italy.
authors: S
authors: S
authors: Rivetti
authors: M
authors: M
authors: Lauriola
authors: M
authors: M
authors: Voltattorni
authors: M
authors: M
authors: Bianchini
authors: D
authors: D
authors: Martini
authors: C
authors: C
authors: Ceccarelli
authors: A
authors: A
authors: Palmieri
authors: G
authors: G
authors: Mattei
authors: M
authors: M
authors: Franchi
authors: G
authors: G
authors: Ugolini
authors: G
authors: G
authors: Rosati
authors: I
authors: I
authors: Montroni
authors: M
authors: M
authors: Taffurelli
authors: Rossella
authors: R
authors: Solmi
mesh_terms: D001426:Bacterial Proteins / Q000494:pharmacology*; D002453:Cell Cycle / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D003110:Colonic Neoplasms / Q000235:genetics*; D004396:Coloring Agents; D018076:DNA, Complementary / Q000096:biosynthesis / Q000235:genetics; D020895:Enterocytes / Q000187:drug effects / Q000378:metabolism; D066246:ErbB Receptors / Q000096:biosynthesis / Q000235:genetics; D005434:Flow Cytometry; D020869:Gene Expression Profiling / Q000379:methods*; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects*; D019073:HT29 Cells; D066257:Heparin-binding EGF-like Growth Factor; D006801:Humans; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism; D046228:Microarray Analysis; D008855:Microscopy, Electron, Scanning; D012334:RNA, Neoplasm / Q000096:biosynthesis / Q000235:genetics; D066247:Receptor, ErbB-4; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014343:Trypan Blue
chemical_list: D001426:Bacterial Proteins; D004396:Coloring Agents; D018076:DNA, Complementary; C587883:HBEGF protein, human; C587884:Hbegf protein, mouse; D066257:Heparin-binding EGF-like Growth Factor; D036341:Intercellular Signaling Peptides and Proteins; D012334:RNA, Neoplasm; C028581:CRM197 (non-toxic variant of diphtheria toxin); C587689:ERBB4 protein, human; D066246:ErbB Receptors; C587690:Erbb4 protein, mouse; D066247:Receptor, ErbB-4; D014343:Trypan Blue