miR-200c is aberrantly expressed in leiomyomas in an ethnic-dependent manner and targets ZEBs, VEGFA, TIMP2, and FBLN5

Abstract

MicroRNA-200c (miR-200c) through repression of specific target genes has been associated with cellular transition, tumorigenesis, and tissue fibrosis. We explored the expression and functional aspects of miR-200c in genesis of leiomyomas (LYO), benign uterine tumors with fibrotic characteristic. Using LYO and matched myometrium (MYO; n=76) from untreated and from patients exposed to hormonal therapies (GNRH agonist (GNRHa), Depo-Provera, and oral contraceptives), we found that miR-200c was expressed at significantly lower levels (P<0.05) in LYO as compared with MYO. These levels were lower in LYO from African Americans as compared with Caucasians, patients experiencing abnormal uterine bleeding and those exposed to GNRHa therapy. Gain-of-function of miR-200c in isolated leiomyoma smooth muscle cells (LSMCs), myometrial smooth muscle cells (MSMCs), and leiomyosarcoma cell line (SKLM-S1) repressed ZEB1/ZEB2 mRNAs and proteins, with concurrent increase in E-cadherin (CDH1) and reduction in vimentin expression, phenotypic alteration, and inhibition of MSMC and LSMC proliferations. We further validated TIMP2, FBLN5, and VEGFA as direct targets of miR-200c through interaction with their respective 3′ UTRs, and other genes as determined by microarray analysis. At tissue levels, LYO expressed lower levels of TIMP2 and FBLN5 mRNAs but increased protein expressions, which to some extent altered due to hormonal exposure. Given the regulatory functions of ZEBs, VEGFA, FBLN5, and TIMP2 on cellular activities that promote cellular transition, angiogenesis, and matrix remodeling, we concluded that altered expression of miR-200c may have a significant impact on the outcome of LYO growth, maintenance of their mesenchymal and fibrotic characteristics, and possibly their associated symptoms.

Effects of miRNA-197 overexpression on proliferation, apoptosis and migration in levonorgestrel treated uterine leiomyoma cells

“…Accumulated evidence suggests that microRNAs (miRNA), a member of non-protein coding small RNA, functions as key regulator of protein coding genes expression [28,29], and their deregulation or dysfunction has been associated with the inflammation, fibrosis and tumorigenesis [30,31]. Several miRNAs, including miR-29b [32], miR-93/106b [33], and miR-200c [34,35], are down-regulated in leiomyoma and involved in the development of uterus leiomyoma. In our study, we found that miR-197 was also down-regulated in human uterus leiomyomas in comparison with the adjacent normal uterus myometrium.…”

“…MiR-93/106b directly and/or indirectly interacted with the 3 0 UTR of F3, IL8, CTGF, PAI-1 and play key roles in myometrium inflammation and tissue turnover [33]. MiR-200c interacted with the 3 0 UTR of IL-8 and targets a potential mediator of inflammation IKBKB in NF-kB signaling pathway [35], miR-200c also interacted with the 3 0 UTR of ZEBs, VEGFA, FBLN5, TIMP2 and promoted cellular transition, angiogenesis, and matrix remodeling [34].…”

The Mechanism and Function of Epigenetics in Uterine Leiomyoma Development

“…Furthermore, several other miRNA direct targets have been characterized in leiomyoma. 82,83,84 More recently, Chuang et al 85 reported that miR-200c regulates IL8 expression by direct targeting IKBKB and alteration of NF-kB activity in leiomyoma.…”

Identification and Verification of Candidate Genes Regulating Neural Stem Cells Behavior Under Hypoxia

“…Subsequently, we performed a search in PubMed for these 14 miRNAs and found that 7 of them had been confirmed experimentally to target VEGFA, that is, miR-361 [40, 41], miR-140 [42], miR-15a [43], miR-15b [44], miR-205 [45], miR-206 [46], and miR-200c [47]. Therefore, we speculate that these miRNAs may target VEGFA and play a role in NSC behavior under hypoxia.…”