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Letrozole Decreased Testosterone-Induced Cell Proliferation and Prolactin Secretion also Increased Apoptosis in MMQ and GH3 Rat Prolactinoma Cell Lines

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Abstract

Aromatase enzyme plays an essential role in estrogen-induced tumorigenesis. It is expressed in the normal pituitary and more significantly in prolactinoma tissues. The aim of this study was to investigate the effects of an aromatase inhibitor, letrozole, on MMQ and GH3 rat prolactinoma cell lines and evaluate the possible mechanism of action. MMQ and GH3 cells were characterized with demonstrating aromatase enzyme and estrogen receptor alpha expression by PCR and immunofluorescence staining. After dose optimization for testosterone (T) and letrozole (L), four groups were established: only the testosteron-treated group (T) to detect cell proliferation; only letrozole-treated group (L) to investigate apoptotic effects; testosterone and letrozole concomitant-treated group to demonstrate inhibition of testosterone induced cell proliferation with letrozole treatment s(T + L) and control group (C) with no treatment. The proliferation rate of cells was determined by WST-1. For the detection of apoptotic and necrotic cells, Annexin V and caspase-3 labeling was used. Prolactin and estrogen levels were measured with ELISA, and the mRNA expression of aromatase and Esr1 was also determined. Testosterone induced the proliferation of MMQ and GH3 cells and further increased prolactin and estradiol levels. Adding letrozole to testosterone resulted in decreased cellular proliferation and even induced apoptosis. Also, letrozole administration significantly decreased prolactin and estradiol levels. However, letrozole alone had no effects on proliferation and apoptosis. Gene expression of aromatase and Esr1 was also significantly decreased by letrozole treatment. This in vitro study demonstrated that treatment of testosterone proliferating cells with letrozole resulted in decreased prolactin levels and cell proliferation and induced apoptosis, and further loss of aromatase and Esr1 mRNA expression were observed. Although this is an in vivo study, the results showed unique and novel findings which may easily be adapted to clinical use for further verification.

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Data Availability

The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

Special thanks is given to Novo-Nordisk for collaboration.

Funding

The study was supported by the Research Fund of the University and Novo-Nordisk collaboration, project no. 2013/155.

Author information

Authors and Affiliations

  1. Department of Endocrinology and Metabolism, Faculty of Medicine, Kocaeli University, Umuttepe Kocaeli, Turkey

    Alev Selek, Berrin Cetinarslan, Zeynep Canturk & Ilhan Tarkun

  2. Department of Medical Biology, Faculty of Medicine, Center for Stem Cell and Gene Therapies Research and Practice, Kocaeli University, Kocaeli University, Umuttepe Kocaeli, Turkey

    Zehra Seda Unal Halbutoğulları

  3. The Department of Bioengineering, Bursa Technical University, Bursa, Turkey

    Çiğdem İnci Aydemir

  4. Department of Medical Biology, Faculty of Medicine, Sakarya University, Sakarya, Turkey

    Gülay Erman

  5. Stembio Stem Cell Tecnologies, Istanbul, Turkey

    Cansu Subaşı

  6. Center for Stem Cell and Tissue Engineering Research & Pracitce, İstinye University, İstanbul, Turkey

    Karaöz Erdal

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  1. Alev Selek
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Contributions

Alev Selek had the idea for the article and drafted it, performed the literature search, data analysis, and writing process, and also revised the work. Zehra Seda Unal Halbutoğulları had literature search, data analysis, and writing process. Çiğdem İnci Aydemir had literature search, main cell culture, apopitosis studies, and writing process. Berrin Cetinarslan had literature search, data interpretation, and discussion. Zeynep antürk had literature search, data interpretation, and discussion. İlhan Tarkun had literature search, data interpretation, and discussion. Gülay Erman, had literature search, ELISA studies, and writing process. Cansu Subaşı had literature search, PCR studies, and writing process. Erdal Karaöz had literature search, data interpretation, and discussion and also revision.

Corresponding author

Correspondence to Alev Selek.

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Selek, A., Halbutoğulları, Z.S.U., Aydemir, Ç.İ. et al. Letrozole Decreased Testosterone-Induced Cell Proliferation and Prolactin Secretion also Increased Apoptosis in MMQ and GH3 Rat Prolactinoma Cell Lines. Mol Neurobiol 60, 2442–2454 (2023). https://doi.org/10.1007/s12035-023-03220-2

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