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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

Time course curve for Gly-Pro-p-nitroanilide hydrolysis in adherent cells monolayer.Results are mean values ± SD for three experiments. *Indicates statistical significance when cervical cancer cell lines were compared to the non-tumorigenic cell line, HaCaT (ANOVA followed by Tukey’s test, p ≤ 0.05).
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pone.0134305.g001: Time course curve for Gly-Pro-p-nitroanilide hydrolysis in adherent cells monolayer.Results are mean values ± SD for three experiments. *Indicates statistical significance when cervical cancer cell lines were compared to the non-tumorigenic cell line, HaCaT (ANOVA followed by Tukey’s test, p ≤ 0.05).

Mentions: In this study, we first investigated the linearity of enzyme activity as a function of time. We used a new approach, which evaluates the DPPIV/CD26 enzymatic activity in intact cells, not in cell lysate, as most of the studies describes. So, in order to investigate the activities related to enzyme anchored onto the cell membrane, we evaluated Gly-Pro-p-nitroanilide hydrolysis in the intact monolayer of adherent cells. The human cervical carcinoma and keratinocyte cell lines were able to promoted Gly-Pro-p-nitroanilide hydrolysis in a linear way, for up to 120 min (Fig 1). In view of these results, to ensure the linearity of the reaction, for enzymatic activity inhibition assays we defined a 60 min incubation time. Furthermore, hydrolysis activities were higher in SiHa and HaCaT cell lines, when compared to HeLa and C33A (Fig 1).


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)

Time course curve for Gly-Pro-p-nitroanilide hydrolysis in adherent cells monolayer.Results are mean values ± SD for three experiments. *Indicates statistical significance when cervical cancer cell lines were compared to the non-tumorigenic cell line, HaCaT (ANOVA followed by Tukey’s test, p ≤ 0.05).
© Copyright Policy
Related In: Results  -  Collection

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

pone.0134305.g001: Time course curve for Gly-Pro-p-nitroanilide hydrolysis in adherent cells monolayer.Results are mean values ± SD for three experiments. *Indicates statistical significance when cervical cancer cell lines were compared to the non-tumorigenic cell line, HaCaT (ANOVA followed by Tukey’s test, p ≤ 0.05).
Mentions: In this study, we first investigated the linearity of enzyme activity as a function of time. We used a new approach, which evaluates the DPPIV/CD26 enzymatic activity in intact cells, not in cell lysate, as most of the studies describes. So, in order to investigate the activities related to enzyme anchored onto the cell membrane, we evaluated Gly-Pro-p-nitroanilide hydrolysis in the intact monolayer of adherent cells. The human cervical carcinoma and keratinocyte cell lines were able to promoted Gly-Pro-p-nitroanilide hydrolysis in a linear way, for up to 120 min (Fig 1). In view of these results, to ensure the linearity of the reaction, for enzymatic activity inhibition assays we defined a 60 min incubation time. Furthermore, hydrolysis activities were higher in SiHa and HaCaT cell lines, when compared to HeLa and C33A (Fig 1).

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