LncRNA H19-Derived miR-675-5p Accelerates the Invasion of Extravillous Trophoblast Cells by Inhibiting GATA2 and Subsequently Activating Matrix Metalloproteinases

Abstract

The invasion of extravillous trophoblast (EVT) cells into the maternal decidua, which plays a crucial role in the establishment of a successful pregnancy, is highly orchestrated by a complex array of regulatory mechanisms. Non-coding RNAs (ncRNAs) that fine-tune gene expression at epigenetic, transcriptional, and post-transcriptional levels are involved in the regulatory mechanisms of EVT cell invasion. However, little is known about the characteristic features of EVT-associated ncRNAs. To elucidate the gene expression profiles of both coding and non-coding transcripts (i.e., mRNAs, long non-coding RNAs (lncRNAs), and microRNAs (miRNAs)) expressed in EVT cells, we performed RNA sequencing analysis of EVT cells isolated from first-trimester placentae. RNA sequencing analysis demonstrated that the lncRNA <i>H19</i> and its derived miRNA <i>miR-675-5p</i> were enriched in EVT cells. Although <i>miR-675-5p</i> acts as a placental/trophoblast growth suppressor, there is little information on the involvement of <i>miR-675-5p</i> in trophoblast cell invasion. Next, we evaluated a possible role of <i>miR-675-5p</i> in EVT cell invasion using the EVT cell lines HTR-8/SVneo and HChEpC1b; overexpression of <i>miR-675-5p</i> significantly promoted the invasion of both EVT cell lines. The transcription factor gene <i>GATA2</i> was shown to be a target of <i>miR-675-5p</i>; moreover, small interfering RNA-mediated <i>GATA2</i> knockdown significantly promoted cell invasion. Furthermore, we identified MMP13 and MMP14 as downstream effectors of <i>miR-675-5p</i>/<i>GATA2</i>-dependent EVT cell invasion. These findings suggest that <i>miR-675-5p</i>-mediated <i>GATA2</i> inhibition accelerates EVT cell invasion by upregulating matrix metalloproteinases.