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Introduction of Somatic Mutation in MED12 Induces Wnt4/β-Catenin and Disrupts Autophagy in Human Uterine Myometrial Cell

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Abstract

Uterine fibroids (UFs) or leiomyoma are frequently associated with somatic mutations in the mediator complex subunit 12 (MED12) gene; however, the function of these mutations in human UF biology is yet to be determined. Herein, we determined the functional role of the most common MED12 somatic mutation in the modulation of oncogenic Wnt4/β-catenin and mammalian target of rapamycin (mTOR) signaling pathways. Using an immortalized human uterine myometrial smooth muscle cell line (UtSM), we constitutively overexpressed either MED12-Wild Type or the most common MED12 somatic mutation (c.131G>A), and the effects of this MED12 mutation were compared between these cell lines. This immortalized cell line was used as a model because it expresses wild type MED12 protein and do not possess MED12 somatic mutations. By comparing the effect between MED12-WT and MED12-mutant (mut) stable cell populations, we observed increased levels of protein expression of Wnt4 and β-catenin in MED12-mut cells as compared with MED12-WT cells. MED12-mut cells also expressed increased levels of mTOR protein and oncogenic cyclin D1 which are hallmarks of cell growth and tumorigenicity. This somatic mutation in MED12 showed an effect on cell-cycle progression by induction of S-phase cells. MED12-mut cells also showed inhibition of autophagy as compared with MED12-WT cells. Together, these findings indicate that the MED12 somatic mutation has the potentials for myometrial cell transformation by dysregulating oncogenic Wnt4/β-catenin and its downstream mTOR signaling which might be associated with autophagy abrogation, cell proliferation, and tumorigenicity.

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Acknowledgments

We would like to thank Ms. Archana Laknaur, research assistant, for ordering lab reagents.

Funding

This study was supported by Augusta University Start up package and by the Research Centers in Minority Institutions (RCMI) pilot grant 2G12RR003032-26 (to SKH), and National Institute of Health (NIH)/R01 grant 2R01HD046228-11 (to AAH).

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Authors and Affiliations

  1. Department of Obstetrics and Gynecology, UIC College of Medicine, Chicago, IL, 60612, USA

    Abdeljabar El Andaloussi & Ayman Al-Hendy

  2. Department of Pathology, UIC College of Medicine, Chicago, IL, 60612, USA

    Nahed Ismail

  3. Department of Molecular Medicine, Institute of Biotechnology, University of Texas Health Science Center at San Antonio, San Antonio, TX, 78229-3900, USA

    Thomas G. Boyer

  4. Department of Otolaryngology-Head & Neck Surgery, Augusta University, Medical College of Georgia, 1120 15th Street, BP 4110, Augusta, GA, 30912, USA

    Sunil K. Halder

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  1. Abdeljabar El Andaloussi
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Correspondence to Sunil K. Halder.

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The authors declare that they have no conflicts of interest.

Additional information

A Precis: Somatic mutations in MED12 can potentially induce oncogenic Wnt4/β-catenin pathways and inhibit cellular autophagy leading to transformation of human normal uterine myometrial cells.

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El Andaloussi, A., Al-Hendy, A., Ismail, N. et al. Introduction of Somatic Mutation in MED12 Induces Wnt4/β-Catenin and Disrupts Autophagy in Human Uterine Myometrial Cell. Reprod. Sci. 27, 823–832 (2020). https://doi.org/10.1007/s43032-019-00084-7

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  • DOI: https://doi.org/10.1007/s43032-019-00084-7

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