MiR-150-5p May Contribute to Pathogenesis of Human Leiomyoma via Regulation of the Akt/p27Kip1 Pathway In Vitro

Abstract

Uterine leiomyoma is found in ~50–80% of women of a reproductive age and is the most common reason for hysterectomy. Recently, posttranscriptional gene silencing by microRNAs (miRs) has been reported as a mechanism for regulating gene expression stability in the pathogenesis of uterine leiomyomas. In this study, miR microarray analysis of leiomyomas and paired myometrial tissue revealed numerous aberrantly expressed miRs, including miR-150. In functional assays, transfection with miR-150 mimic resulted in decreased migration and fibrosis, implying an inhibition of leiomyoma growth. To identify the target genes of miR-150 in leiomyoma, gene set analysis and network analysis were performed. To overcome the limitations of in silico analysis, changes in expression levels of hallmark genes in leiomyoma after transfection with a miR-150 mimic were also evaluated using qRT-PCR. As a result, the Akt/p27Kip1 pathway was presumed to be one of the target pathways of miR-150. After transfecting cultured leiomyoma cells with the miR-150 mimic, expression levels of its target gene Akt decreased, whereas those of p27Kip1 increased significantly. Our results suggest that miR-150 affects the cell cycle regulation in uterine leiomyoma through the Akt/p27Kip1 pathway.

MicroRNA-139-5p Regulates Fibrotic Potentials via Modulation of Collagen Type 1 and Phosphorylated p38 MAPK in Uterine Leiomyoma

“…We utilized a previously published method to culture leiomyoma and myometrial smooth muscle cells. 16 Tissue samples were cut into small pieces (~2–5 mm 3 ) and incubated in Dulbecco's modified Eagle's medium without phenol red (Sigma-Aldrich, St. Louis, MO, USA), containing collagenase type I 2.0 mg/mL (Gibco, Waltham, MA, USA), 1% antibiotic-antimycotic mixture containing 100 IU/mL penicillin and 100 mg/mL streptomycin, and 10% heat-inactivated fetal bovine serum (FBS) for 45 min at 37℃ in a shaker. The digested tissue was subsequently cultured using the explant method in a humidified incubator at 37℃ and 5% CO 2 for 2–4 h. Cells were routinely passaged using Versene-EDTA (Gibco).…”

“…Many of these miRNAs are associated with biological pathways related to uterine leiomyomas, including tumorigenesis (let-7 and miR-146), angiogenesis (miR-15 and miR-16), cell cycle regulation (miR-21, miR-150, miR-181, miR-182, and miR-197), inflammation (miR-93, miR-106, and miR-200), and extracellular matrix (ECM) accumulation (miR-21 and miR-29). 13 14 15 16 17 18 19 20 …”

“…Our microarray analysis showed that miR-139-5p was most significantly downregulated among aberrantly expressed 250 miRs. 16 However, its function and mechanism in uterine leiomyomas remain unknown. Therefore, this study aimed to determine the functional role of miR-139-5p in the pathophysiology of uterine leiomyomas.…”

The Role of miRNA and Related Pathways in Pathophysiology of Uterine Fibroids—From Bench to Bedside

“…These authors have found that after transfecting the cells the expression levels of Akt decreased, whereas p27Kip1 significantly increased. Hence, there is a strong evidence that miR-150 may affect cell cycle regulation in UFs by interaction with the Akt-signaling pathway [91]. Since PI3K/Akt/mTOR is an important intracellular signaling pathway in both normal cell cycle regulation and in UFs development the future research could be directed towards unraveling more details of the described interactions [21].…”

MicroRNA‑150 functions as a tumor suppressor and sensitizes osteosarcoma to doxorubicin‑induced apoptosis by targeting RUNX2

“…miRNA-150 (miR-150) is a tumor-associated miRNA, which has been reported as a biomarker in several different types of cancer, including osteosarcoma (15), gastric cancer (16), non-small cell lung cancer (17). Colorectal cancer (18) and leiomyoma (19). However, the biological function and the underlying mechanisms of miR-150 in OS have not yet been investigated.…”