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, Sunil K Halder

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

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<0.05) reduced levels of Wnt4 and β-catenin proteins as well as cell proliferation, as compared to scrambled control cells. Med12 knockdown cells also showed reduced levels of cell-cycle associated cyclin D1, Cdk1, and Cdk2 proteins as well as reduced activation of p-ERK, p-AKT, and TGFβ signaling pathways as compared to scrambled control cells. Moreover, TGFβ regulated fibrosis-related proteins such as fibronectin, collagen type 1, and PAI-1 were significantly (p<0.05) reduced in Med12 knockdown cells as compared to scrambled control cells. Together, these results suggest that Med12 play a key role in the regulation of HuLM cell proliferation through the modulation of Wnt/β-catenin, cell-cycle associated, and fibrosis-associated protein expression.

Original languageEnglish (US)
Pages (from-to)en20161097
JournalEndocrinology
DOIs
StateE-pub ahead of print - Dec 14 2016
Externally publishedYes

Fingerprint

Catenins
Wnt Signaling Pathway
Leiomyoma
Gene Silencing
Lentivirus
Cell Cycle
Proteins
Fibrosis
Cell Proliferation
Wnt4 Protein
Small Interfering RNA
Mediator Complex
Steroid Receptors
Cyclin D1
Plasminogen Activator Inhibitor 1
Collagen Type I
RNA Interference
Fibronectins
Genes
Fluorescence

Cite this

Silencing Med12 Gene Reduces Proliferation of Human Leiomyoma Cells Mediated via Wnt/β-Catenin Signaling Pathway. / Al-Hendy, Ayman; Laknaur, Archana; Diamond, Michael P; Ismail, Nahed; Boyer, Thomas G; Halder, Sunil K.

In: Endocrinology, 14.12.2016, p. en20161097.

Research output: Contribution to journalArticle

Al-Hendy, Ayman ; Laknaur, Archana ; Diamond, Michael P ; Ismail, Nahed ; Boyer, Thomas G ; Halder, Sunil K. / Silencing Med12 Gene Reduces Proliferation of Human Leiomyoma Cells Mediated via Wnt/β-Catenin Signaling Pathway. In: Endocrinology. 2016 ; pp. en20161097.
@article{65a2364f60814659803b5f7d940d7e9e,
title = "Silencing Med12 Gene Reduces Proliferation of Human Leiomyoma Cells Mediated via Wnt/β-Catenin Signaling Pathway",
abstract = "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<0.05) reduced levels of Wnt4 and β-catenin proteins as well as cell proliferation, as compared to scrambled control cells. Med12 knockdown cells also showed reduced levels of cell-cycle associated cyclin D1, Cdk1, and Cdk2 proteins as well as reduced activation of p-ERK, p-AKT, and TGFβ signaling pathways as compared to scrambled control cells. Moreover, TGFβ regulated fibrosis-related proteins such as fibronectin, collagen type 1, and PAI-1 were significantly (p<0.05) reduced in Med12 knockdown cells as compared to scrambled control cells. Together, these results suggest that Med12 play a key role in the regulation of HuLM cell proliferation through the modulation of Wnt/β-catenin, cell-cycle associated, and fibrosis-associated protein expression.",
author = "Ayman Al-Hendy and Archana Laknaur and Diamond, {Michael P} and Nahed Ismail and Boyer, {Thomas G} and Halder, {Sunil K}",
year = "2016",
month = "12",
day = "14",
doi = "10.1210/en.2016-1097",
language = "English (US)",
pages = "en20161097",
journal = "Endocrinology",
issn = "0013-7227",
publisher = "The Endocrine Society",

}

TY - JOUR

T1 - Silencing Med12 Gene Reduces Proliferation of Human Leiomyoma Cells Mediated via Wnt/β-Catenin Signaling Pathway

AU - Al-Hendy, Ayman

AU - Laknaur, Archana

AU - Diamond, Michael P

AU - Ismail, Nahed

AU - Boyer, Thomas G

AU - Halder, Sunil K

PY - 2016/12/14

Y1 - 2016/12/14

N2 - 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<0.05) reduced levels of Wnt4 and β-catenin proteins as well as cell proliferation, as compared to scrambled control cells. Med12 knockdown cells also showed reduced levels of cell-cycle associated cyclin D1, Cdk1, and Cdk2 proteins as well as reduced activation of p-ERK, p-AKT, and TGFβ signaling pathways as compared to scrambled control cells. Moreover, TGFβ regulated fibrosis-related proteins such as fibronectin, collagen type 1, and PAI-1 were significantly (p<0.05) reduced in Med12 knockdown cells as compared to scrambled control cells. Together, these results suggest that Med12 play a key role in the regulation of HuLM cell proliferation through the modulation of Wnt/β-catenin, cell-cycle associated, and fibrosis-associated protein expression.

AB - 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<0.05) reduced levels of Wnt4 and β-catenin proteins as well as cell proliferation, as compared to scrambled control cells. Med12 knockdown cells also showed reduced levels of cell-cycle associated cyclin D1, Cdk1, and Cdk2 proteins as well as reduced activation of p-ERK, p-AKT, and TGFβ signaling pathways as compared to scrambled control cells. Moreover, TGFβ regulated fibrosis-related proteins such as fibronectin, collagen type 1, and PAI-1 were significantly (p<0.05) reduced in Med12 knockdown cells as compared to scrambled control cells. Together, these results suggest that Med12 play a key role in the regulation of HuLM cell proliferation through the modulation of Wnt/β-catenin, cell-cycle associated, and fibrosis-associated protein expression.

U2 - 10.1210/en.2016-1097

DO - 10.1210/en.2016-1097

M3 - Article

C2 - 27967206

SP - en20161097

JO - Endocrinology

JF - Endocrinology

SN - 0013-7227

ER -