Limits...
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

DPPIV/CD26 expression profile determined by real-time RT-PCR analysis.Quantitative real-time RT-PCR analysis of DPPIV/CD26 expression in cervical cancer cells (SiHa, HeLa and C33A) and immortalized keratinocytes (HaCaT). The total RNA was isolated and the cDNA was analyzed with the primers as described in Materials and methods section. Bars represent mean ± 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
getmorefigures.php?uid=PMC4519168&req=5

pone.0134305.g002: DPPIV/CD26 expression profile determined by real-time RT-PCR analysis.Quantitative real-time RT-PCR analysis of DPPIV/CD26 expression in cervical cancer cells (SiHa, HeLa and C33A) and immortalized keratinocytes (HaCaT). The total RNA was isolated and the cDNA was analyzed with the primers as described in Materials and methods section. Bars represent mean ± 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: To better investigate if the cervical cancer cells and HaCaT could present difference in the DPPIV/CD26 gene and protein expression, we performed quantitative real-time RT-PCR and flow cytometry analysis. The result showed that mRNA and protein levels of expression (Figs 2 and 3) correlates with the enzyme activity presented in the adherent cervical cancer cells and HaCaT (Fig 1). As observed for the enzymatic profile of adherent cells, HaCaT and SiHa presented the highest DPPIV/CD26 expression. However, in the C33A cell line this protein was expressed at low levels, while for the HeLa cell line, the DPPIV/CD26 was almost undetectable (Figs 2 and 3).


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)

DPPIV/CD26 expression profile determined by real-time RT-PCR analysis.Quantitative real-time RT-PCR analysis of DPPIV/CD26 expression in cervical cancer cells (SiHa, HeLa and C33A) and immortalized keratinocytes (HaCaT). The total RNA was isolated and the cDNA was analyzed with the primers as described in Materials and methods section. Bars represent mean ± 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.g002: DPPIV/CD26 expression profile determined by real-time RT-PCR analysis.Quantitative real-time RT-PCR analysis of DPPIV/CD26 expression in cervical cancer cells (SiHa, HeLa and C33A) and immortalized keratinocytes (HaCaT). The total RNA was isolated and the cDNA was analyzed with the primers as described in Materials and methods section. Bars represent mean ± 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: To better investigate if the cervical cancer cells and HaCaT could present difference in the DPPIV/CD26 gene and protein expression, we performed quantitative real-time RT-PCR and flow cytometry analysis. The result showed that mRNA and protein levels of expression (Figs 2 and 3) correlates with the enzyme activity presented in the adherent cervical cancer cells and HaCaT (Fig 1). As observed for the enzymatic profile of adherent cells, HaCaT and SiHa presented the highest DPPIV/CD26 expression. However, in the C33A cell line this protein was expressed at low levels, while for the HeLa cell line, the DPPIV/CD26 was almost undetectable (Figs 2 and 3).

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