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Differential Expression and Enzymatic Activity of DPPIV/CD26 Affects Migration Ability of Cervical Carcinoma Cells.

Beckenkamp A, Willig JB, Santana DB, Nascimento J, Paccez JD, Zerbini LF, Bruno AN, Pilger DA, Wink MR, Buffon A - PLoS ONE (2015)

Bottom Line: This enzyme is found mainly anchored onto the cell membrane, although it also has a soluble form, an enzymatically active isoform.We observed higher migratory capacity of HeLa, when compared to SiHa.However this mechanism seems to be mediated independent of DPPIV/CD26.

View Article: PubMed Central - PubMed

Affiliation: Pharmaceutical Sciences Graduate Program, Faculty of Pharmacy, Federal University of Rio Grande do Sul (UFRGS), Porto Alegre, RS, Brazil.

ABSTRACT
Dipeptidyl peptidase IV (DPPIV/CD26) is a transmembrane glycoprotein that inactivates or degrades some bioactive peptides and chemokines. For this reason, it regulates cell proliferation, migration and adhesion, showing its role in cancer processes. This enzyme is found mainly anchored onto the cell membrane, although it also has a soluble form, an enzymatically active isoform. In the present study, we investigated DPPIV/CD26 activity and expression in cervical cancer cell lines (SiHa, HeLa and C33A) and non-tumorigenic HaCaT cells. The effect of the DPPIV/CD26 inhibitor (sitagliptin phosphate) on cell migration and adhesion was also evaluated. Cervical cancer cells and keratinocytes exhibited DPPIV/CD26 enzymatic activity both membrane-bound and in soluble form. DPPIV/CD26 expression was observed in HaCaT, SiHa and C33A, while in HeLa cells it was almost undetectable. We observed higher migratory capacity of HeLa, when compared to SiHa. But in the presence of sitagliptin SiHa showed an increase in migration, indicating that, at least in part, cell migration is regulated by DPPIV/CD26 activity. Furthermore, in the presence of sitagliptin phosphate, SiHa and HeLa cells exhibited a significant reduction in adhesion. However this mechanism seems to be mediated independent of DPPIV/CD26. This study demonstrates, for the first time, the activity and expression of DPPIV/CD26 in cervical cancer cells and the effect of sitagliptin phosphate on cell migration and adhesion.

No MeSH data available.


Related in: MedlinePlus

Effect of sitagliptin phosphate on cell adhesion.Comparison of the adhesion on plastic plates uncoated or coated with ECM proteins (laminin, fibronectin, type I collagen and type IV collagen), of cervical cancer cells SiHa (a) and HeLa (b) 2 h after plating in the absence (control) and presence of 0.2 or 1 mM sitagliptin phosphate. Data were presented as the ratio of treatment absorbance/control absorbance. Results are mean values ± SD (n = 3). *Indicates statistical significance when sitagliptin phosphate groups were compared to the respective control group. (ANOVA followed by Tukey’s test, p ≤ 0.05).
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pone.0134305.g005: Effect of sitagliptin phosphate on cell adhesion.Comparison of the adhesion on plastic plates uncoated or coated with ECM proteins (laminin, fibronectin, type I collagen and type IV collagen), of cervical cancer cells SiHa (a) and HeLa (b) 2 h after plating in the absence (control) and presence of 0.2 or 1 mM sitagliptin phosphate. Data were presented as the ratio of treatment absorbance/control absorbance. Results are mean values ± SD (n = 3). *Indicates statistical significance when sitagliptin phosphate groups were compared to the respective control group. (ANOVA followed by Tukey’s test, p ≤ 0.05).

Mentions: In order to determine whether cell adhesion of SiHa and HeLa cell lines could be affected by the DPPIV/CD26 inhibitor, sitagliptin phosphate, we plated these cells in the presence or absence of the inhibitor and cell adhesion was assessed after 2 h of incubation. With this incubation time, control cells adhere on plastic, without spreading. But in the presence of ECM proteins (laminin, fibronectin and type I collagen), after 2 hours has been observed cell spreading and increased adhesion capacity (S4 Fig). The results indicated that after 2 h, HeLa cell line adheres more than SiHa. Moreover, it was observed that, in the presence of sitagliptin phosphate 1 mM, both cell lines exhibit a significant reduction in cell adhesion in plastic plates uncoated or coated with ECM proteins (Fig 5). Whereas sitagliptin phosphate has affected adhesion also in HeLa cell line, which practically does not express DPPIV/CD26, we believe that this effect may be mediated independent of DPPIV/CD26 enzymatic activity.


Differential Expression and Enzymatic Activity of DPPIV/CD26 Affects Migration Ability of Cervical Carcinoma Cells.

Beckenkamp A, Willig JB, Santana DB, Nascimento J, Paccez JD, Zerbini LF, Bruno AN, Pilger DA, Wink MR, Buffon A - PLoS ONE (2015)

Effect of sitagliptin phosphate on cell adhesion.Comparison of the adhesion on plastic plates uncoated or coated with ECM proteins (laminin, fibronectin, type I collagen and type IV collagen), of cervical cancer cells SiHa (a) and HeLa (b) 2 h after plating in the absence (control) and presence of 0.2 or 1 mM sitagliptin phosphate. Data were presented as the ratio of treatment absorbance/control absorbance. Results are mean values ± SD (n = 3). *Indicates statistical significance when sitagliptin phosphate groups were compared to the respective control group. (ANOVA followed by Tukey’s test, p ≤ 0.05).
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4519168&req=5

pone.0134305.g005: Effect of sitagliptin phosphate on cell adhesion.Comparison of the adhesion on plastic plates uncoated or coated with ECM proteins (laminin, fibronectin, type I collagen and type IV collagen), of cervical cancer cells SiHa (a) and HeLa (b) 2 h after plating in the absence (control) and presence of 0.2 or 1 mM sitagliptin phosphate. Data were presented as the ratio of treatment absorbance/control absorbance. Results are mean values ± SD (n = 3). *Indicates statistical significance when sitagliptin phosphate groups were compared to the respective control group. (ANOVA followed by Tukey’s test, p ≤ 0.05).
Mentions: In order to determine whether cell adhesion of SiHa and HeLa cell lines could be affected by the DPPIV/CD26 inhibitor, sitagliptin phosphate, we plated these cells in the presence or absence of the inhibitor and cell adhesion was assessed after 2 h of incubation. With this incubation time, control cells adhere on plastic, without spreading. But in the presence of ECM proteins (laminin, fibronectin and type I collagen), after 2 hours has been observed cell spreading and increased adhesion capacity (S4 Fig). The results indicated that after 2 h, HeLa cell line adheres more than SiHa. Moreover, it was observed that, in the presence of sitagliptin phosphate 1 mM, both cell lines exhibit a significant reduction in cell adhesion in plastic plates uncoated or coated with ECM proteins (Fig 5). Whereas sitagliptin phosphate has affected adhesion also in HeLa cell line, which practically does not express DPPIV/CD26, we believe that this effect may be mediated independent of DPPIV/CD26 enzymatic activity.

Bottom Line: This enzyme is found mainly anchored onto the cell membrane, although it also has a soluble form, an enzymatically active isoform.We observed higher migratory capacity of HeLa, when compared to SiHa.However this mechanism seems to be mediated independent of DPPIV/CD26.

View Article: PubMed Central - PubMed

Affiliation: Pharmaceutical Sciences Graduate Program, Faculty of Pharmacy, Federal University of Rio Grande do Sul (UFRGS), Porto Alegre, RS, Brazil.

ABSTRACT
Dipeptidyl peptidase IV (DPPIV/CD26) is a transmembrane glycoprotein that inactivates or degrades some bioactive peptides and chemokines. For this reason, it regulates cell proliferation, migration and adhesion, showing its role in cancer processes. This enzyme is found mainly anchored onto the cell membrane, although it also has a soluble form, an enzymatically active isoform. In the present study, we investigated DPPIV/CD26 activity and expression in cervical cancer cell lines (SiHa, HeLa and C33A) and non-tumorigenic HaCaT cells. The effect of the DPPIV/CD26 inhibitor (sitagliptin phosphate) on cell migration and adhesion was also evaluated. Cervical cancer cells and keratinocytes exhibited DPPIV/CD26 enzymatic activity both membrane-bound and in soluble form. DPPIV/CD26 expression was observed in HaCaT, SiHa and C33A, while in HeLa cells it was almost undetectable. We observed higher migratory capacity of HeLa, when compared to SiHa. But in the presence of sitagliptin SiHa showed an increase in migration, indicating that, at least in part, cell migration is regulated by DPPIV/CD26 activity. Furthermore, in the presence of sitagliptin phosphate, SiHa and HeLa cells exhibited a significant reduction in adhesion. However this mechanism seems to be mediated independent of DPPIV/CD26. This study demonstrates, for the first time, the activity and expression of DPPIV/CD26 in cervical cancer cells and the effect of sitagliptin phosphate on cell migration and adhesion.

No MeSH data available.


Related in: MedlinePlus