Z Geburtshilfe Neonatol 2011; 215 - PO11_08
DOI: 10.1055/s-0031-1293422

MicroRNA expression profiles in trophoblastic cells

DM Morales Prieto 1, M Weber 2, S Ospina-Prieto 1, S Hölters 1, E Schleußner 2, UR Markert 2
  • 1Universitätsklinikum Jena, Klinik für Frauenheilkunde und Geburtshilfe, Abteilung Geburtshilfe, Jena
  • 2Universitätsklinikum Jena, Frauenklinik, Abt. Geburtshilfe, Jena

Ziel: MicroRNAs are small single-stranded RNA molecules playing an important role in the post-transcriptional regulation of gene expression. Aim of this study was to analyze miRNA expression in four trophoblastic cell lines (JEG–3, ACH–3P, AC1-M59, HTR8/SV neo) before and after LIF challenge.

Methodik: Cells were stimulated or not with LIF for 4h and total RNA containing microRNAs was isolated. Expression of 762 microRNAs was screened in biological duplicates. HTR–8 cells were karyotyped through Multiplex-Fluorescence In Situ Hybridisation (M-FISH). Furthermore, expression kinetics of miR–9, miR–141, miR–21, miR–93 and let–7g has been analyzed by real time PCR in JEG–3 after stimulation with LIF for 1 to 24h. As being the most affected, miR–141 has been silenced and over-expressed to test its role in proliferation of JEG–3 cells after 24h and 48h.

Ergebnis: MicroRNAs expression profiles of JEG–3, ACH–3P and AC1-M59 were similar exhibing high expression of the C19MC miRNA cluster, known to be almost exclusively expressed by the placenta, as well as the three miRNAs recognized as specific markers for human embryonic stem cells. Conversely, HTR–8 exhibits almost no expression of the C19MC miRNAs but instead high expression of a miRNA cluster located in the chromosome 14. These results correlate with those observed in the M-FISH karyotype of HTR–8 cells, where both C19 and C14 were found to be physically altered. MicroRNAs change differentially between cell lines under LIF stimulation. Only a small group of miRNAs were found simultaneously altered in all tested cells. Silencing of miR–141 completely inhibited proliferation of JEG–3 cells, while overexpression had no effect.

Schlussfolgerung: LIF modulates microRNA expression in trophoblastic cells in a cell type-dependent manner. Karyotype and miRNA expression profile provide a tool for the identification of trophoblast cells and their malignant derivates. MiR–141 may have an essential role in regulation of functions in trophoblastic cells.