Abstract
BackgroundHuman uterine leiomyoma (fibroids; LYO) are the most common benign neoplasms in reproductive-aged women. Dysregulated extracellular matrix and irregular LYO reversion-inducing cysteine-rich protein with Kazal motifs (RECK) expression are thought to be mediated by aberrant microRNA (miR) expression. The relationship of miR-15b and RECK expression in LYO has not been studied.MethodsThe expression levels of miR-15b and RECK were determined by quantitative RT-PCR, Western blot, and immunohistochemistry in cultures derived from commercial primary leiomyoma (cpLYO) and myometrial (cpMYO) cell lines and leiomyoma (pLYO) and myometrium (pMYO) tissue from surgical samples respectively. The relationship between miR-15b and RECK expression in cpLYO and pLYO (compared to their respective myometrial controls) was evaluated following transfection of cell cultures with either miR-15b mimic or inhibitor.ResultsElevated levels of miR-15b were observed in cpLYO (2.82-fold; p = 0.04) and pLYO cell (1.30-fold; p = 0.0001) cultures respectively compared to corresponding MYO cell controls. Following transfection with miR-15b mimic, cpLYO cells (0.62-fold; p < 0.0001) and pLYO cells (0.68-fold; p < 0.0001) demonstrated reduced RECK protein expression. Following transfection with miR-15b inhibitor, cpLYO cells (1.20-fold; p < 0.0001) and pLYO cells (1.31-fold; p = 0.0007) demonstrated elevated RECK protein expression. RECK protein expression was reduced in pLYO tissues (0.73-fold; p < 0.0001) and pLYO (0.47-fold; p = 0.047) cells when compared to the corresponding MYO tissue controls.ConclusionOur findings suggest that miR-15b negatively regulates RECK expression in LYO, and increased miR-15b and decreased RECK expression may contribute to the pathobiology of LYO. The functional significance of miR-15b and RECK expression warrants further investigation as potential therapeutic targets for the treatment of human LYO.
Highlights
Human uterine leiomyoma are the most common benign neoplasms in reproductive-aged women
Increased expression of miR-15b in human Human uterine leiomyoma (LYO) MiR-15b was observed to be differentially expressed between human leiomyoma and matched myometrial tissues in our miRNA microarray analysis
MiR-15b was increased in commercial primary leiomyoma (cpLYO) cell line (2.82-fold; p = 0.04, Fig. 1b) and Primary leiomyoma (pLYO) cells (1.3-fold; p < 0.0001, Fig. 1c) when compared with their MYO cell controls, respectively
Summary
Human uterine leiomyoma (fibroids; LYO) are the most common benign neoplasms in reproductive-aged women. Human uterine leiomyoma (fibroids; LYO) are benign uterine smooth muscle tumors with an estimated overall incidence of 75 % among reproductive-aged women, and are symptomatic in approximately 25 % of women [1,2,3]. This gynecologic disorder may result in heavy and painful menstrual bleeding, pelvic pressure and pain, recurrent pregnancy loss, and infertility. Differential microRNA (miRNA) expression has been implicated as a possible contributor to the pathobiology of LYO [9]
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