RNA干扰INHα基因对绵羊颗粒细胞周期及凋亡相关基因表达的影响

doi:10.11843/j.issn.0366-6964.2017.08.021

Abstract

Abstract:

The aim of this study was to investigate the effects of silencing the inhibin α-subunit (INHα) gene by RNAi on cell cycle and apoptosis-related genes in sheep granulosa cells (GCs), explore the local regulation of INHα on sheep GCs, which can provide a foundation for improving the livestock's fecundity. siINHα and siNC (negative control) were transfected into GCs from antral follicles (3-7 mm) obtained from the ovaries of Thin-tailed Han sheep (1.0-1.5 years old). Non-transfected cells were as blank control (Control). The mRNA expression of INHα and the genes related to cell cycle and apoptosis were analyzed by quantitative real-time PCR (qRT-PCR). The cell cycle progression was measured by flow cytometry. The results showed that siRNA was transfected into sheep GCs at 48 h with the silencing efficiency over 85%. Knockdown of INHα by siRNA significantly decreased the mRNA expressions of the cell cycle related genes CyclinD1 and Cyclin B1 by 0.2 and 0.7 times, respectively (P<0.05), but increased the mRNA expression of p21 by 0.9 times (P<0.05). Meanwhile, the mRNA expressions of pro-apoptotic genes Bax, p53 and Caspase3 were increased by 1.8, 3.6 and 0.4 times, respectively (P<0.05), but no significant changes of anti-apoptosis gene Bcl-2 compared with negative control. There was no significant difference between the control and negative control (P>0.05). INHα gene silencing altered the progression of cell cycle as well. The percentage of cell at S phase was decreased, while the percentage of cell at G1 phase was significantly increased compared with the negative control (P<0.05). No significant difference was observed between control and negative control (P>0.05). These results indicated that INHα was a key regulator in GCs and involved in the regulation of folliculogenesis by regulating the growth and apoptosis of GCs in sheep.

CLC Number:

S826.2

LI Ting, MA Ai-tuan, ZHANG Ying-jie, LIU Yue-qin, DUAN Chun-hui. Effects of Silencing INHα Gene by RNAi on Cycle,Apoptosis-Related Genes in Sheep Granulosa Cells[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(8): 1551-1556.

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Effects of Silencing INHα Gene by RNAi on Cycle,Apoptosis-Related Genes in Sheep Granulosa Cells

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