Effects of Silencing Gpr3 Gene by Small Interfering RNA on Apoptosis in Porcine Granulosa Cells

Abstract: G protein-coupled receptor 3 (Gpr3)， a member of the G protein-coupled receptor superfamily， is known as a critical factor for the maintenance of meiotic prophase arrest in oocytes via a Gs protein-mediated pathway， but its role in granulosa cells is still unclear. In this study， RNAi was used to explore the effect of silencing Gpr3 gene expression on apoptosis of porcine granulosa cells (GCs). The specific siRNAs against Gpr3 was designed， chemically synthesized and transiently transfected into the procine GCs using lipidosome. Gpr3 expression level was assayed by Real-time PCR and Western blot. Cytoactive， apoptotic and the expression level of apoptotic-related genes were analyzed by MTT， flow cytometry and Real-time PCR， respectively. The results showed that Gpr3-siRNA effectively inhibited the expression of Gpr3 at both mRNA and protein levels in porcine GCs (P<0.01); the cytoactivity of porcine GCs increased from 0.419 to 0.586， and the apoptosis rate dropped from 2.67% to 0.42%; the expression levels of Bcl-2 were significantly increased， while the expression levels of Bax were reduced (P<0.05). Our results suggested that siRNA-mediated Gpr3 depletion inhibited apoptosis in porcine granulosa cells， which were possibly related to the expression levels of Bcl-2 and Bax genes.

CSTR: 32200.14.cjcb.2014.08.0011