title: Mechanisms underlying aberrant expression of miR-29c in uterine leiomyoma.
abstract: OBJECTIVE
  To determine the expression of miR-29c and its target genes in leiomyoma and the role of NF-κB, specific protein 1 (SP1), and DNA methylation in its regulation.
  
  
  METHODS
  Experimental study.
  
  
  METHODS
  Academic research laboratory.
  
  
  METHODS
  Women undergoing hysterectomy for leiomyoma.
  
  
  METHODS
  Over- and underexpression of miR-29c; blockade of transcription factors.
  
  
  METHODS
  MiR-29c and its target gene levels in leiomyoma and the effects of blockade of transcription factors on miR-29c expression.
  
  
  RESULTS
  Leiomyoma as compared with myometrium expressed significantly lower levels of miR-29c, with an inverse relationship with expression of its targets, COL3A1 and DNMT3A. Gain of function of miR-29c inhibited the expression of COL3A1 and DNMT3A at protein and mRNA levels, secreted COL3A1, and rate of cell proliferation. Loss of function of miR-29c had the opposite effect. E2, P, and their combination inhibited miR-29c in leiomyoma smooth muscle cells (LSMC). Phosphorylated NF-κB (p65) and SP1 protein expression were significantly increased in leiomyoma. SiRNA knockdown of SP1 and DNMT3A or their specific inhibitors significantly increased the expression of miR-29c, accompanied by the inhibition of cellular and secreted COL3A1 in siRNA-treated cells. Knockdown of p65 also induced miR-29c expression but had no effect on COL3A1 expression.
  
  
  CONCLUSIONS
  MiR-29c expression is suppressed in leiomyoma, resulting in an increase in expression of its targets COL3A1 and DNMT3A. The suppression of miR-29c in LSMC is primarily mediated by SP1, NF-κB signaling, and epigenetic modification. Collectively, these results indicate a significant role for miR-29c in leiomyoma pathogenesis.
authors.affiliation: Department of Obstetrics and Gynecology, Harbor-UCLA Medical Center and LA-Biomed Research Institute, Torrance, California.
authors.forename: Tsai-Der
authors.initials: TD
authors.lastname: Chuang
authors.affiliation: Department of Obstetrics and Gynecology, Harbor-UCLA Medical Center and LA-Biomed Research Institute, Torrance, California. Electronic address: okhorram@obgyn.humc.edu.
authors.forename: Omid
authors.initials: O
authors.lastname: Khorram
mesh_terms: D000328:Adult; D049109:Cell Proliferation; D024061:Collagen Type III / Q000235:genetics / Q000378:metabolism; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000235:genetics / Q000378:metabolism; D019175:DNA Methylation; D000090205:DNA Methyltransferase 3A; D015536:Down-Regulation; D044127:Epigenesis, Genetic; D004958:Estradiol / Q000494:pharmacology; D005260:Female; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D007889:Leiomyoma / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D008875:Middle Aged; D010766:Phosphorylation; D011374:Progesterone / Q000494:pharmacology; D034622:RNA Interference; D016329:Sp1 Transcription Factor / Q000235:genetics / Q000378:metabolism; D051996:Transcription Factor RelA / Q000378:metabolism; D014158:Transcription, Genetic; D014162:Transfection; D014407:Tumor Cells, Cultured; D014594:Uterine Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology
chemical_list: C439971:COL3A1 protein, human; D024061:Collagen Type III; C000717967:DNMT3A protein, human; C532045:MIRN29a microRNA, human; D035683:MicroRNAs; C498143:RELA protein, human; D016329:Sp1 Transcription Factor; C108209:SP1 protein, human; D051996:Transcription Factor RelA; D011374:Progesterone; D004958:Estradiol; D004248:DNA (Cytosine-5-)-Methyltransferases; D000090205:DNA Methyltransferase 3A
keywords: COL3A1; DNMT3A; Leiomyoma; SP1; miR-29c