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PKN1 modulates TGFβ and EGF signaling in HEC-1-A endometrial cancer cell line.

Attarha S, Saini RK, Andersson S, Mints M, Souchelnytskyi S - Onco Targets Ther (2014)

Bottom Line: The response of cells to TGFβ and EGF is mediated by a network of various intracellular regulators.It was observed that phosphorylation of Smad2, FAK, and Erk1/2 correlated with responses of the cells to TGFβ1 and EGF.PKN1 modulates TGFβ- and EGF-dependent regulation of cell proliferation, migration, and invasiveness, and therefore is a component of the network signaling downstream of TGFβ and EGF.

View Article: PubMed Central - PubMed

Affiliation: Department of Oncology-Pathology, Karolinska Institutet, Stockholm, Sweden ; Department of Women's and Children's Health, Karolinska Institutet, Stockholm, Sweden.

ABSTRACT

Background: The response of cells to TGFβ and EGF is mediated by a network of various intracellular regulators. The signaling crosstalk between different regulators is of key importance for tumorigenesis. The crosstalk may explain the modulation of cellular responses to the same regulator by another signaling molecule. As PKN1 - a serine/threonine kinase implicated in tumorigenesis - was identified as potential crosstalk node for TGFβ and EGF signaling, the cellular functions that may be affected by PKN1 in a crosstalk of TGFβ and EGF were explored.

Methods: To investigate the contribution of PKN1 to TGFβ and EGF signaling, transiently PKN1-transfected HEC-1-A endometrial cancer cells were generated and subjected to treatment with TGFβ1, EGF, and their combination. Proliferation, apoptosis, invasion, wound healing, and migration assays were performed. The impact of PKN1 on the expression and phosphorylation of intracellular proteins was monitored by immunoblotting.

Results: It was demonstrated that PKN1 modulated the responses of HEC-A-1 endometrial cancer cells to TGFβ1 and EGF. PKN1 had an inhibitory effect on the stimulation of cell migration, and PKN1 kinase activity was required for the inhibitory effect of TGFβ and EGF on cell proliferation and invasiveness. It was observed that phosphorylation of Smad2, FAK, and Erk1/2 correlated with responses of the cells to TGFβ1 and EGF.

Conclusion: PKN1 modulates TGFβ- and EGF-dependent regulation of cell proliferation, migration, and invasiveness, and therefore is a component of the network signaling downstream of TGFβ and EGF.

No MeSH data available.


Related in: MedlinePlus

PKN1 is a potential component of TGFβ and EGF crosstalk.Notes: (A) The expression of WT, KN, and CA PKN1 constructs upon transfection in HEC-1-A cells. The migration position of PKN1 constructs is indicated. (B) The network for PKN1 proposed in the context of the authors’ previous proteomics study17 is shown. Links between components define functional interactions.Abbreviations: CA, constitutively active; KN, kinase negative; WT, wild type.
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f1-ott-7-1397: PKN1 is a potential component of TGFβ and EGF crosstalk.Notes: (A) The expression of WT, KN, and CA PKN1 constructs upon transfection in HEC-1-A cells. The migration position of PKN1 constructs is indicated. (B) The network for PKN1 proposed in the context of the authors’ previous proteomics study17 is shown. Links between components define functional interactions.Abbreviations: CA, constitutively active; KN, kinase negative; WT, wild type.

Mentions: PKN1 has been identified as a protein deregulated in endometrial cancer.17 To study the impact of PKN1 on cells, WT, KN, and CA PKN1 was transiently expressed in HEC-1-A endometrial cancer cells. The expression of PKN1 was controlled by immunoblotting with anti-PKN1 antibody (Figure 1A). Expression of endogenous PKN1 was observed in six out of seven tested cell lines (Figure S1). Enhanced expression of PKN1 constructs allowed the impact of PKN1 on cell physiology to be accentuated (Figure 1A). Transiently transfected cells were then used in the tests described below. Systemic analysis of potential connections between PKN1, TGFβ, and EGF showed involvement of a number of potent regulators of cell proliferation and cytoskeleton rearrangement (Figure 1B). This prompted the exploration of whether PKN1 plays a role in modulation of TGFβ- and EGF-dependent regulation of cell proliferation, migration, and invasiveness.


PKN1 modulates TGFβ and EGF signaling in HEC-1-A endometrial cancer cell line.

Attarha S, Saini RK, Andersson S, Mints M, Souchelnytskyi S - Onco Targets Ther (2014)

PKN1 is a potential component of TGFβ and EGF crosstalk.Notes: (A) The expression of WT, KN, and CA PKN1 constructs upon transfection in HEC-1-A cells. The migration position of PKN1 constructs is indicated. (B) The network for PKN1 proposed in the context of the authors’ previous proteomics study17 is shown. Links between components define functional interactions.Abbreviations: CA, constitutively active; KN, kinase negative; WT, wild type.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4128851&req=5

f1-ott-7-1397: PKN1 is a potential component of TGFβ and EGF crosstalk.Notes: (A) The expression of WT, KN, and CA PKN1 constructs upon transfection in HEC-1-A cells. The migration position of PKN1 constructs is indicated. (B) The network for PKN1 proposed in the context of the authors’ previous proteomics study17 is shown. Links between components define functional interactions.Abbreviations: CA, constitutively active; KN, kinase negative; WT, wild type.
Mentions: PKN1 has been identified as a protein deregulated in endometrial cancer.17 To study the impact of PKN1 on cells, WT, KN, and CA PKN1 was transiently expressed in HEC-1-A endometrial cancer cells. The expression of PKN1 was controlled by immunoblotting with anti-PKN1 antibody (Figure 1A). Expression of endogenous PKN1 was observed in six out of seven tested cell lines (Figure S1). Enhanced expression of PKN1 constructs allowed the impact of PKN1 on cell physiology to be accentuated (Figure 1A). Transiently transfected cells were then used in the tests described below. Systemic analysis of potential connections between PKN1, TGFβ, and EGF showed involvement of a number of potent regulators of cell proliferation and cytoskeleton rearrangement (Figure 1B). This prompted the exploration of whether PKN1 plays a role in modulation of TGFβ- and EGF-dependent regulation of cell proliferation, migration, and invasiveness.

Bottom Line: The response of cells to TGFβ and EGF is mediated by a network of various intracellular regulators.It was observed that phosphorylation of Smad2, FAK, and Erk1/2 correlated with responses of the cells to TGFβ1 and EGF.PKN1 modulates TGFβ- and EGF-dependent regulation of cell proliferation, migration, and invasiveness, and therefore is a component of the network signaling downstream of TGFβ and EGF.

View Article: PubMed Central - PubMed

Affiliation: Department of Oncology-Pathology, Karolinska Institutet, Stockholm, Sweden ; Department of Women's and Children's Health, Karolinska Institutet, Stockholm, Sweden.

ABSTRACT

Background: The response of cells to TGFβ and EGF is mediated by a network of various intracellular regulators. The signaling crosstalk between different regulators is of key importance for tumorigenesis. The crosstalk may explain the modulation of cellular responses to the same regulator by another signaling molecule. As PKN1 - a serine/threonine kinase implicated in tumorigenesis - was identified as potential crosstalk node for TGFβ and EGF signaling, the cellular functions that may be affected by PKN1 in a crosstalk of TGFβ and EGF were explored.

Methods: To investigate the contribution of PKN1 to TGFβ and EGF signaling, transiently PKN1-transfected HEC-1-A endometrial cancer cells were generated and subjected to treatment with TGFβ1, EGF, and their combination. Proliferation, apoptosis, invasion, wound healing, and migration assays were performed. The impact of PKN1 on the expression and phosphorylation of intracellular proteins was monitored by immunoblotting.

Results: It was demonstrated that PKN1 modulated the responses of HEC-A-1 endometrial cancer cells to TGFβ1 and EGF. PKN1 had an inhibitory effect on the stimulation of cell migration, and PKN1 kinase activity was required for the inhibitory effect of TGFβ and EGF on cell proliferation and invasiveness. It was observed that phosphorylation of Smad2, FAK, and Erk1/2 correlated with responses of the cells to TGFβ1 and EGF.

Conclusion: PKN1 modulates TGFβ- and EGF-dependent regulation of cell proliferation, migration, and invasiveness, and therefore is a component of the network signaling downstream of TGFβ and EGF.

No MeSH data available.


Related in: MedlinePlus