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Expression of Nodal, Cripto, SMAD3, Phosphorylated SMAD3, and SMAD4 in the Proliferative Endometrium of Women With Endometriosis

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Abstract

Background

Nodal is a growth factor of the transforming growth factor β superfamily that is expressed in high turnover tissues, such as the human endometrium, and in several malignancies. The effects of Nodal are modulated by the coreceptor Cripto and mediated by SMAD proteins. This study evaluated the gene and protein expression of Nodal, Cripto, total and phosphorylated (p) SMAD3, and SMAD4 in the proliferative endometrium of women with and without endometriosis.

Method

Total RNA was isolated and complementary DNA synthesized from eutopic endometrium of women with (n = 15) and without (n = 12) endometriosis, followed by quantitative real-time polymerase chain reaction (PCR) to evaluate the gene expression of Nodal, Cripto, SMAD3, and SMAD4. Western blot was used to evaluate the protein levels of Nodal and Cripto, and immunohistochemistry was performed to localize SMAD3, pSMAD3, and SMAD4.

Results

Although Nodal expression was unchanged in women with endometriosis, real-time PCR indicated lower gene expression of Cripto (fold change 0.27, P < .05) in the endometriosis group. This difference, however, was not maintained at protein expression level as assessed by Western blot. The immunostaining of total SMAD3 was reduced in the endometriosis group (P < .01), but the localization of pSMAD3 and the nuclear staining of SMAD4 were unchanged.

Conclusion

These findings suggest that the Nodal signaling pathway has subtle changes in the endometrium of women with endometriosis, but this imbalance may not cause functional damage as it seems not to affect the nuclear expression of SMAD4.

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Author information

Authors and Affiliations

  1. Division of Human Reproduction, Department of Obstetrics and Gynecology, Federal University of Minas Gerais, Av. Alfredo Balena, 110, 9° andar, 30130-100, Belo Horizonte, Minas Gerais, Brazil

    Cynthia Dela Cruz PhD, Helen L. Del Puerto PhD, Ana Luiza L. Rocha MD, PhD, Inês K. Cavallo MD, Alessandra D. Clarizia MD, PhD & Fernando M. Reis MD, PhD

  2. Department of Molecular and Developmental Medicine, Section of Obstetrics and Gynecology, University of Siena, Siena, Italy

    Felice Petraglia MD, FRCOG

Authors
  1. Cynthia Dela Cruz PhD
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  2. Helen L. Del Puerto PhD
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  3. Ana Luiza L. Rocha MD, PhD
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  4. Inês K. Cavallo MD
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  5. Alessandra D. Clarizia MD, PhD
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  6. Felice Petraglia MD, FRCOG
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  7. Fernando M. Reis MD, PhD
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Corresponding author

Correspondence to Fernando M. Reis MD, PhD.

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Cruz, C.D., Del Puerto, H.L., Rocha, A.L.L. et al. Expression of Nodal, Cripto, SMAD3, Phosphorylated SMAD3, and SMAD4 in the Proliferative Endometrium of Women With Endometriosis. Reprod. Sci. 22, 527–533 (2015). https://doi.org/10.1177/1933719114549855

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