Silencing Med12 Gene Reduces Proliferation of Human Leiomyoma Cells Mediated via Wnt/β-Catenin Signaling Pathway Ayman Al-Hendy, Archana Laknaur, Michael P. Diamond, Nahed Ismail, Thomas G. Boyer, and Sunil K. Halder Endocrinology Endocrine Society Submitted: February 11, 2016 Accepted: December 08, 2016 First Online: December 14, 2016   Early Release articles are PDF versions of manuscripts that have been peer reviewed and accepted  but not yet copyedited. The manuscripts are published online as soon as possible after acceptance  and before the copyedited, typeset articles are published. They are posted "as is" (i.e., as  submitted by the authors at the modification stage), and do not reflect editorial changes. No  corrections/changes to the PDF manuscripts are accepted. Accordingly, there likely will be  differences between the Early Release manuscripts and the final, typeset articles. The manuscripts  remain listed on the Early Release page until the final, typeset articles are posted. At that point,  the manuscripts are removed from the Early Release page.    DISCLAIMER: These manuscripts are provided "as is" without warranty of any kind, either express  or particular purpose, or non‐infringement. Changes will be made to these manuscripts before  publication. Review and/or use or reliance on these materials is at the discretion and risk of the  reader/user. In no event shall the Endocrine Society be liable for damages of any kind arising  references to, products or publications do not imply endorsement of that product or publication. 

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Endocrinology; Copyright 2016

DOI: 10.1210/en.2016-1097

Silencing Med12 Gene Reduces Proliferation of Human Leiomyoma Cells Mediated via Wnt/β-Catenin Signaling Pathway Ayman Al-Hendy1, Archana Laknaur1, Michael P. Diamond1, Nahed Ismail2, Thomas G. Boyer3, and Sunil K. Halder 1* 1

Department of Obstetrics and Gynecology, Augusta University, Medical College of Georgia, Georgia 30912, United States; 2Clinical Microbiology Division, University of Pittsburgh, Pittsburgh, PA, 15261, United States; 3 Department of Molecular Medicine, Institute of Biotechnology, University of Texas Health Science Center at San Antonio, San Antonio, Texas 78229-3900, United States

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Received 11 February 2016. Accepted 8 December 2016. Effect of Med12 silencing on HuLM cell growth

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Abbreviations: Mediator complex subunit 12; Med12; uterine fibroids; leiomyoma; fibrosis; TGFβ3;

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Uterine fibroids, or leiomyoma, are the most common benign tumors in women of reproductive age. Herein, the effect of silencing the mediator complex subunit 12 (Med12) gene in human uterine fibroid cells was evaluated. The role of Med12 in the modulation of Wnt/β-catenin and cell-proliferation-associated signaling was evaluated in human uterine fibroid cells. Med12 was silenced in the immortalized human uterine fibroid cell line (HuLM) using a lentivirus-based Med12 gene-specific RNA interference (RNAi) strategy. HuLM cells were infected with lentiviruses carrying Med12-specific short shRNA sequences or a non-functional shRNA scrambled control with green fluorescence protein (GFP). Stable cells that expressed low levels of Med12 protein were characterized. Wnt/β-catenin signaling, sex steroid receptor signaling, cell-cycle associated, and fibrosis associated proteins were measured. Med12 knockdown cells showed significantly (p