Involvement of miR-105-1, miR-152, Vinculin and α-Parvin in the Control of Porcine Ovarian Cell Functions. Do miRNAs and siRNAs Need a Transfection Reagent?

The aim of the present study was to examine the role of miR-105-1, miR-152, vinculin (VCL) and α-parvin (PARVA) in the control of ovarian cell functions, and whether miRNAs and siRNAs could enter and affect cultured cells with and without the application of carriers. For this purpose, we (1) examined the incorporation, expression and effect of the selected miRNAs (miR-105-1 and miR-152 mimics and antagonists) and siRNAs downregulating VCL and PARVA on cultured porcine ovarian granulosa cells; and (2) compared the incorporation, expression, and effects of these miRNAs and siRNAs used without and with carriers (transfection reagent: Lipofectamine). The incorporation and expression of these RNAs, viability, proliferation, apoptosis and steroid hormone release of cells were analysed by immunocytochemistry, RT-qPCR, the alamarBlue test, BrdU incorporation, a DNA fragmentation (Cell Death Detection) kit and ELISA. The obtained results demonstrate (1) that miR-105-1, miR-152, VCL and PARVA can be stimulators of the main basic ovarian cell functions; (2) that miRNAs can be incorporated and expressed in ovarian cells and affect their functions without a carrier; and (3) that a carrier is not necessary for siRNA incorporation and expression but is necessary for their influence on ovarian cell functions.