Gene expression changes and promoter methylation with the combined effects of estradiol and leptin in uterine tissue of the ovariectomized mice model of menopause

  • Abhishek Shetty
  • Thejaswini Venkatesh
  • Rie Tsutsumi
  • Padmanaban S. SureshEmail author
Original Article


Substantial epidemiological studies have shown an association of obesity with the common gynecological malignancy, endometrial cancer. The relevant interactions and contribution of estradiol and the adipose cytokine, leptin, in endometrial lesions are not completely understood. Suitable animal models to understand the physiological response of uterine tissue to the combined effects of estradiol–leptin are lacking. To investigate the effect of estradiol–leptin crosstalk on gene expression and associated altered pathways, we established an ovariectomized mouse model, treated with 17-β estradiol (0.1 µg/mouse subcutaenously., for every 12 h) and/or recombinant mouse leptin (1 μg/g Bwt intraperitoneally., for every 12 h) for 4 h, 20 h, and 40 h. Gene expressions by semi-quantitative RT-PCR, uterine tissue protein phosphorylation status by western blotting and promoter methylation were analyzed in estradiol, progesterone insufficient animals. Semi-quantitative RT-PCR demonstrated significantly increased expression of Esr, Igf1, Igfbp3, Vegfr1, and Vegf, and significantly decreased expression of Mmp9 after co-treatment with estradiol and leptin, indicating a common transcriptional network regulated by the treatments. Ovariectomy-induced histomorphological changes were only reversed by estradiol. Methylation-specific PCR, analyzing methylation of CpG sites of Vegfa, Pgr, and Igf1, revealed that transcriptional regulation after hormonal treatments is independent of methylation at the examined CpG sites. Western blot confirmed the increased expression of PSTAT-3 (Ser-727) and PERK1/2 proteins after estradiol + leptin treatment, confirming the estradiol + leptin cross-talk hypothesis. In conclusion, our in vivo studies determined specific gene expression and signaling protein changes, and further unraveled the molecular targets of estradiol + leptin that may perturb endometrial homeostasis and lead to endometrial hyperplasia development in the chronic stimulated state.


Uterine tissue Endocrine physiology Hormone response 



PSS acknowledges Department of science and technology- Science engineering research board (YSS/2014/000020), India for the grant support. PSS also acknowledge University Grants Commission, India for start up-grant. We are also grateful to the Department of Science and Technology-Fund for Improvement of Science and Technology (DST-FIST) and University Grants Commission Special assistance program for providing infrastructure support to the Department of Biosciences, Mangalore University. PSS was affiliated with Mangalore University earlier and acknowledge the support of the university for the conduct of the work.

Compliance with ethical standards

Conflict of interest

On behalf of all authors, the corresponding author states that there is no conflict of interest.

Ethical approval

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.


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Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Department of BiosciencesMangalore UniversityMangaloreIndia
  2. 2.Department of Biochemistry and Molecular BiologyCentral University of KeralaKasaragodIndia
  3. 3.Department of Nutrition and Metabolism. Institute of Biomedical SciencesTokushima University Graduate SchoolTokushima CityJapan
  4. 4.School of BiotechnologyNational Institute of TechnologyCalicutIndia

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