Gene expression profiles of miR-100-5p-transfected normal endometrial stromal cells

Accumulating evidence suggests that microRNAs play definite roles in the pathogenesis of endometriosis. The objective of the study was to determine the role of miR-100-5p, one of the upregulated microRNA in endometriotic cyst stromal cells, in the pathogenesis of endometriosis. Downstream targets of miR-100-5p were identified by compulsory expression of miR-100-5p in normal eutopic endometrial stromal cells (NESCs), a global mRNA microarray technique, and pathways analysis.Compulsory expression of miR-100-5p in NESCs directed the induction of cell motility through SWItch/sucrose non-fermentable (SWI/SNF)-related matrix-associated actin-dependent regulator of chromatin subfamily D member 1 (SMARCD1) supression and matrix metallopeptidase 1 (MMP1) activation. NESCs were transfected with precursor hsa-miR-100-5p (Pre-miRTM miRNA precursor- hsa-miR-100, Ambion, Austin, TX, USA) or negative control precursor miRNA (Pre-miRTM miRNA precursor-negative control #1 Ambion) at a final concentration of 10 nM, using LipofectamineTM RNAiMAX (Invitrogen, Carlsbad, CA, USA). Forty-eight hours after transfection, total RNA from cultured NESCs transfected with precursor hsa-miR-100-5p (n=4) and NESCs transfected with negative control precursor miRNA (n=4) was extracted with an RNeasy Mini kit (Qiagen, Valencia, CA, USA). Then, the samples were subjected to a gene expression microarray analysis with a commercially available human mRNA microarray (G4851A, SurePrint G3 Human Gene Expression Microarray 8x60K v2, Agilent Technologies, Santa Clara, CA, USA).