Limits...
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.

Show MeSH

Related in: MedlinePlus

Invasion ability evaluation on exendin-4 treated cells.Representative experiment from three independent experiments on the invasive ability of SK-N-AS and SH-SY5Y cells after treatment with 0.3 µM exendin-4 (A); real-time RT-PCR analysis of the expression of MMP-9 and TIMP-1 in NB cells after treatment with 0.3 µM exendin-4 for 6 h. * = p<0.05 vs. related control (B). Size of the cell colonies grown in soft agar 7, 14 or 21 days after suspension, with or without (C = control) exendin-4. Data are reported as mean percentage vs. related control of three replicates. EXE = exendin-4; * = p<0.05 vs. related control; # = p<0.05 vs. C 7d; § = p<0.05 vs. C 14d (C).
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3750033&req=5

pone-0071716-g010: Invasion ability evaluation on exendin-4 treated cells.Representative experiment from three independent experiments on the invasive ability of SK-N-AS and SH-SY5Y cells after treatment with 0.3 µM exendin-4 (A); real-time RT-PCR analysis of the expression of MMP-9 and TIMP-1 in NB cells after treatment with 0.3 µM exendin-4 for 6 h. * = p<0.05 vs. related control (B). Size of the cell colonies grown in soft agar 7, 14 or 21 days after suspension, with or without (C = control) exendin-4. Data are reported as mean percentage vs. related control of three replicates. EXE = exendin-4; * = p<0.05 vs. related control; # = p<0.05 vs. C 7d; § = p<0.05 vs. C 14d (C).

Mentions: The invasive potential of cells in the presence of exendin-4 was evaluated using a modified Matrigel Boyden chamber assay. Exendin-4 significantly decreased cell invasion in SK-N-AS. In agreement with this finding, a marked increase in the expression level of TIMP-1 was observed (Fig. 10 A–B). On the contrary, exendin-4 increased cell invasion in SH-SY5Y cells and this effect was paralleled by a significant increase of both MMP-9 and TIMP-1 expression levels (Fig. 10 A–B). However SH-SY5Y, in contrast to SK-N-AS, are also able to grow in soft agar and this anchorage-independent growth is correlated with in vivo oncogenic potential. We performed a long term exendin-4 treatment (7–21 days) of SH-SY5Y cells in soft agar assay and we observed a significant inhibition of the size of the colonies (Fig. 10C). Altogether, these findings suggest that exendin-4 promotes the acquisition of a more differentiated phenotype in SH-SY5Y cells.


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)

Invasion ability evaluation on exendin-4 treated cells.Representative experiment from three independent experiments on the invasive ability of SK-N-AS and SH-SY5Y cells after treatment with 0.3 µM exendin-4 (A); real-time RT-PCR analysis of the expression of MMP-9 and TIMP-1 in NB cells after treatment with 0.3 µM exendin-4 for 6 h. * = p<0.05 vs. related control (B). Size of the cell colonies grown in soft agar 7, 14 or 21 days after suspension, with or without (C = control) exendin-4. Data are reported as mean percentage vs. related control of three replicates. EXE = exendin-4; * = p<0.05 vs. related control; # = p<0.05 vs. C 7d; § = p<0.05 vs. C 14d (C).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3750033&req=5

pone-0071716-g010: Invasion ability evaluation on exendin-4 treated cells.Representative experiment from three independent experiments on the invasive ability of SK-N-AS and SH-SY5Y cells after treatment with 0.3 µM exendin-4 (A); real-time RT-PCR analysis of the expression of MMP-9 and TIMP-1 in NB cells after treatment with 0.3 µM exendin-4 for 6 h. * = p<0.05 vs. related control (B). Size of the cell colonies grown in soft agar 7, 14 or 21 days after suspension, with or without (C = control) exendin-4. Data are reported as mean percentage vs. related control of three replicates. EXE = exendin-4; * = p<0.05 vs. related control; # = p<0.05 vs. C 7d; § = p<0.05 vs. C 14d (C).
Mentions: The invasive potential of cells in the presence of exendin-4 was evaluated using a modified Matrigel Boyden chamber assay. Exendin-4 significantly decreased cell invasion in SK-N-AS. In agreement with this finding, a marked increase in the expression level of TIMP-1 was observed (Fig. 10 A–B). On the contrary, exendin-4 increased cell invasion in SH-SY5Y cells and this effect was paralleled by a significant increase of both MMP-9 and TIMP-1 expression levels (Fig. 10 A–B). However SH-SY5Y, in contrast to SK-N-AS, are also able to grow in soft agar and this anchorage-independent growth is correlated with in vivo oncogenic potential. We performed a long term exendin-4 treatment (7–21 days) of SH-SY5Y cells in soft agar assay and we observed a significant inhibition of the size of the colonies (Fig. 10C). Altogether, these findings suggest that exendin-4 promotes the acquisition of a more differentiated phenotype in SH-SY5Y cells.

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