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01.12.2012 | Research | Ausgabe 1/2012 Open Access

Reproductive Biology and Endocrinology 1/2012

Protein expression of PKCZ (Protein Kinase C Zeta), Munc18c, and Syntaxin-4 in the insulin pathway in endometria of patients with polycystic ovary syndrome (PCOS)

Zeitschrift:
Reproductive Biology and Endocrinology > Ausgabe 1/2012
Autoren:
Rodrigo Rivero, Claire-Alix Garin, Paulina Ormazabal, Andrea Silva, Rodrigo Carvajal, Fernando Gabler, Carmen Romero, Margarita Vega
Wichtige Hinweise

Electronic supplementary material

The online version of this article (doi:10.​1186/​1477-7827-10-17) contains supplementary material, which is available to authorized users.

Competing interests

The authors declare that they have no competing interests.

Authors' contributions

RR, CAG, PO, and MV conceived and designed the experiments. RR, CAG, PO, and AS performed the experiments; RR, CAG, PO, AS, CR and MV analyzed the data. MV and CR contributed reagents/materials/analysis tools. RR, CAG and MV wrote the manuscript. All authors read and approved the final manuscript.

Abstract

Background

Polycystic Ovary Syndrome (PCOS) is an endocrine-metabolic disorder commonly associated with insulin resistance (IR). Previous studies indicate about the expression of molecules involved in the insulin pathway in endometria of women with PCOS-IR. Therefore, the aim of the present study was to evaluate the effect of insulin and testosterone in the expression of these proteins in the endometria and immortal endometrial stromal cell line (T-HESCs).

Methods

We examined the protein levels of Munc18c, PKC zeta, phospho-PKC Zeta, and Syntaxin-4. Protein levels were assessed by Western Blot and/or immunohistochemistry in proliferative endometria (NPE = 6) and in PCOS endometria with insulin resistance (PCOSE-IR = 6). We also evaluated whether high concentrations of insulin (100 nM) and/or testosterone (100 nM), during a 24 h stimulatory period, affected the expression of these proteins in an immortal endometrial stromal cell line (T-HESCs). Once stimulated, proteins were extracted from cells and were assessed by Western Blot analysis. Immunocytochemistry was performed to detect AR in T-HESC cells.

Results

Western Blot data showed decreased expression (p < 0,05) of Munc18c and phospho-PKC Zeta in PCOS-IR endometria (PCOSE-IR) with respect to the control (NPE). In the in vitro study, Western Blot analysis showed decreased levels of Munc18c, PKC Zeta and phospho-PKC Zeta with the different hormonal treatments when compared to the control condition (no hormonal stimulation) (p < 0,05). The AR was present in the endometrial stromal cell line (T-HESC).

Conclusion

The conditions of hyperinsulinism and hyperandrogenism present in PCOS-IR patients modulate the expression and/or phosphorylation of the proteins involved in the insulin pathway at the endometrial level. These data extend to the T-HESCs cells results, where insulin and testosterone exert an effect on both the expression and phosphorylation of proteins present in the pathway.
Zusatzmaterial
Authors’ original file for figure 1
12958_2011_958_MOESM1_ESM.pdf
Authors’ original file for figure 2
12958_2011_958_MOESM2_ESM.pdf
Authors’ original file for figure 3
12958_2011_958_MOESM3_ESM.pdf
Authors’ original file for figure 4
12958_2011_958_MOESM4_ESM.pdf
Literatur
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