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Exendin-4 induces cell adhesion and differentiation and counteracts the invasive potential of human neuroblastoma cells.

Luciani P, Deledda C, Benvenuti S, Squecco R, Cellai I, Fibbi B, Marone IM, Giuliani C, Modi G, Francini F, Vannelli GB, Peri A - PLoS ONE (2013)

Bottom Line: More recently, additional biological properties have been associated to molecules that belong to the GLP-1 family.However, no data are currently available on the effects of exendin-4 on tumor cell motility.Furthermore, we demonstrated that exendin-4 reduced cell migration and counteracted anchorage-independent growth in neuroblastoma cells.

View Article: PubMed Central - PubMed

Affiliation: Endocrine Unit, "Center for Research, Transfer and High Education on Chronic, Inflammatory, Degenerative and Neoplastic Disorders for the Development of Novel Therapies" (DENOThe), Department of Experimental and Clinical Biomedical Sciences, University of Florence, Florence, Italy.

ABSTRACT
Exendin-4 is a molecule currently used, in its synthetic form exenatide, for the treatment of type 2 diabetes mellitus. Exendin-4 binds and activates the Glucagon-Like Peptide-1 Receptor (GLP-1R), thus inducing insulin release. More recently, additional biological properties have been associated to molecules that belong to the GLP-1 family. For instance, Peptide YY and Vasoactive Intestinal Peptide have been found to affect cell adhesion and migration and our previous data have shown a considerable actin cytoskeleton rearrangement after exendin-4 treatment. However, no data are currently available on the effects of exendin-4 on tumor cell motility. The aim of this study was to investigate the effects of this molecule on cell adhesion, differentiation and migration in two neuroblastoma cell lines, SH-SY5Y and SK-N-AS. We first demonstrated, by Extra Cellular Matrix cell adhesion arrays, that exendin-4 increased cell adhesion, in particular on a vitronectin substrate. Subsequently, we found that this molecule induced a more differentiated phenotype, as assessed by i) the evaluation of neurite-like protrusions in 3D cell cultures, ii) the analysis of the expression of neuronal markers and iii) electrophysiological studies. Furthermore, we demonstrated that exendin-4 reduced cell migration and counteracted anchorage-independent growth in neuroblastoma cells. Overall, these data indicate for the first time that exendin-4 may have anti-tumoral properties.

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Analysis of passive properties and ionic current densities in FNC and SK-N-AS cells.Data were evaluated until 48 h in culture. Effect of Exendin-4 (EXE) on RMP (A), on membrane capacitance, Cm (B), and resting specific membrane conductance Gm/Cm (C). * p<0.05 EXE- treated cells vs. the related control; § and §§ p<0.05 and <0.01 SK-N-AS 48 h vs. 24 h; †<0.05 and †† <0.01 SK-N-AS+Exendin-4 48 h vs. 24 h. FNC at 48 h, SK-N-AS at 24 and 48 h data are from 26, 28 and 32 cells, respectively. Effect of EXE on ISAC/Cm (D), INa/Cm (E), ICa,T/Cm (F) and ICa,L/Cm (G). * and ** p<0.05 and P<0.01 EXE treated cells vs. the related control; §, §§ and §§§ p<0.05, <0.01 and <0.001 SK-N-AS 48 h vs. 24 h; †<0.05, †† <0.01 and ††† 0.001 SKNAS +EXE 48 h vs. 24 h. ISAC/Cm INa/Cm, ICa,T/Cm and ICa,L/Cm in FNC at 48 h, SK-N-AS at 24 and 48 h data in each experimental condition are from 14–18 cells.
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pone-0071716-g006: Analysis of passive properties and ionic current densities in FNC and SK-N-AS cells.Data were evaluated until 48 h in culture. Effect of Exendin-4 (EXE) on RMP (A), on membrane capacitance, Cm (B), and resting specific membrane conductance Gm/Cm (C). * p<0.05 EXE- treated cells vs. the related control; § and §§ p<0.05 and <0.01 SK-N-AS 48 h vs. 24 h; †<0.05 and †† <0.01 SK-N-AS+Exendin-4 48 h vs. 24 h. FNC at 48 h, SK-N-AS at 24 and 48 h data are from 26, 28 and 32 cells, respectively. Effect of EXE on ISAC/Cm (D), INa/Cm (E), ICa,T/Cm (F) and ICa,L/Cm (G). * and ** p<0.05 and P<0.01 EXE treated cells vs. the related control; §, §§ and §§§ p<0.05, <0.01 and <0.001 SK-N-AS 48 h vs. 24 h; †<0.05, †† <0.01 and ††† 0.001 SKNAS +EXE 48 h vs. 24 h. ISAC/Cm INa/Cm, ICa,T/Cm and ICa,L/Cm in FNC at 48 h, SK-N-AS at 24 and 48 h data in each experimental condition are from 14–18 cells.

Mentions: When recorded in physiological bath solution the resting membrane potential (RMP) was similar in both control and exendin-4 treated cell models at 48 h, whereas exendin-4-treated SK-N-AS cells appeared more depolarized at 24 h (Fig. 6A). The cell-surface area evaluated by the membrane capacitance (Cm) was significantly higher (Fig. 6B), whereas the specific resting membrane conductance (Gm/Cm) was reduced in both exendin-4-treated SK-N-AS and FNC (Fig. 6C). Such a reduction of Gm/Cm is in agreement with diminished membrane leak currents and is an index of cell differentiation induced by exendin-4. Accordingly, the Gm/Cm values in SK-N-AS at 24 h were greater than those recorded at 48 h in both control and exendin-4-treated cells. The results obtained in SK-N-AS at 48 h were similar to those observed in SH- SY5Y at the same time, as previously reported [17].


Exendin-4 induces cell adhesion and differentiation and counteracts the invasive potential of human neuroblastoma cells.

Luciani P, Deledda C, Benvenuti S, Squecco R, Cellai I, Fibbi B, Marone IM, Giuliani C, Modi G, Francini F, Vannelli GB, Peri A - PLoS ONE (2013)

Analysis of passive properties and ionic current densities in FNC and SK-N-AS cells.Data were evaluated until 48 h in culture. Effect of Exendin-4 (EXE) on RMP (A), on membrane capacitance, Cm (B), and resting specific membrane conductance Gm/Cm (C). * p<0.05 EXE- treated cells vs. the related control; § and §§ p<0.05 and <0.01 SK-N-AS 48 h vs. 24 h; †<0.05 and †† <0.01 SK-N-AS+Exendin-4 48 h vs. 24 h. FNC at 48 h, SK-N-AS at 24 and 48 h data are from 26, 28 and 32 cells, respectively. Effect of EXE on ISAC/Cm (D), INa/Cm (E), ICa,T/Cm (F) and ICa,L/Cm (G). * and ** p<0.05 and P<0.01 EXE treated cells vs. the related control; §, §§ and §§§ p<0.05, <0.01 and <0.001 SK-N-AS 48 h vs. 24 h; †<0.05, †† <0.01 and ††† 0.001 SKNAS +EXE 48 h vs. 24 h. ISAC/Cm INa/Cm, ICa,T/Cm and ICa,L/Cm in FNC at 48 h, SK-N-AS at 24 and 48 h data in each experimental condition are from 14–18 cells.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3750033&req=5

pone-0071716-g006: Analysis of passive properties and ionic current densities in FNC and SK-N-AS cells.Data were evaluated until 48 h in culture. Effect of Exendin-4 (EXE) on RMP (A), on membrane capacitance, Cm (B), and resting specific membrane conductance Gm/Cm (C). * p<0.05 EXE- treated cells vs. the related control; § and §§ p<0.05 and <0.01 SK-N-AS 48 h vs. 24 h; †<0.05 and †† <0.01 SK-N-AS+Exendin-4 48 h vs. 24 h. FNC at 48 h, SK-N-AS at 24 and 48 h data are from 26, 28 and 32 cells, respectively. Effect of EXE on ISAC/Cm (D), INa/Cm (E), ICa,T/Cm (F) and ICa,L/Cm (G). * and ** p<0.05 and P<0.01 EXE treated cells vs. the related control; §, §§ and §§§ p<0.05, <0.01 and <0.001 SK-N-AS 48 h vs. 24 h; †<0.05, †† <0.01 and ††† 0.001 SKNAS +EXE 48 h vs. 24 h. ISAC/Cm INa/Cm, ICa,T/Cm and ICa,L/Cm in FNC at 48 h, SK-N-AS at 24 and 48 h data in each experimental condition are from 14–18 cells.
Mentions: When recorded in physiological bath solution the resting membrane potential (RMP) was similar in both control and exendin-4 treated cell models at 48 h, whereas exendin-4-treated SK-N-AS cells appeared more depolarized at 24 h (Fig. 6A). The cell-surface area evaluated by the membrane capacitance (Cm) was significantly higher (Fig. 6B), whereas the specific resting membrane conductance (Gm/Cm) was reduced in both exendin-4-treated SK-N-AS and FNC (Fig. 6C). Such a reduction of Gm/Cm is in agreement with diminished membrane leak currents and is an index of cell differentiation induced by exendin-4. Accordingly, the Gm/Cm values in SK-N-AS at 24 h were greater than those recorded at 48 h in both control and exendin-4-treated cells. The results obtained in SK-N-AS at 48 h were similar to those observed in SH- SY5Y at the same time, as previously reported [17].

Bottom Line: More recently, additional biological properties have been associated to molecules that belong to the GLP-1 family.However, no data are currently available on the effects of exendin-4 on tumor cell motility.Furthermore, we demonstrated that exendin-4 reduced cell migration and counteracted anchorage-independent growth in neuroblastoma cells.

View Article: PubMed Central - PubMed

Affiliation: Endocrine Unit, "Center for Research, Transfer and High Education on Chronic, Inflammatory, Degenerative and Neoplastic Disorders for the Development of Novel Therapies" (DENOThe), Department of Experimental and Clinical Biomedical Sciences, University of Florence, Florence, Italy.

ABSTRACT
Exendin-4 is a molecule currently used, in its synthetic form exenatide, for the treatment of type 2 diabetes mellitus. Exendin-4 binds and activates the Glucagon-Like Peptide-1 Receptor (GLP-1R), thus inducing insulin release. More recently, additional biological properties have been associated to molecules that belong to the GLP-1 family. For instance, Peptide YY and Vasoactive Intestinal Peptide have been found to affect cell adhesion and migration and our previous data have shown a considerable actin cytoskeleton rearrangement after exendin-4 treatment. However, no data are currently available on the effects of exendin-4 on tumor cell motility. The aim of this study was to investigate the effects of this molecule on cell adhesion, differentiation and migration in two neuroblastoma cell lines, SH-SY5Y and SK-N-AS. We first demonstrated, by Extra Cellular Matrix cell adhesion arrays, that exendin-4 increased cell adhesion, in particular on a vitronectin substrate. Subsequently, we found that this molecule induced a more differentiated phenotype, as assessed by i) the evaluation of neurite-like protrusions in 3D cell cultures, ii) the analysis of the expression of neuronal markers and iii) electrophysiological studies. Furthermore, we demonstrated that exendin-4 reduced cell migration and counteracted anchorage-independent growth in neuroblastoma cells. Overall, these data indicate for the first time that exendin-4 may have anti-tumoral properties.

Show MeSH
Related in: MedlinePlus