FOXO3基因过表达对鹅卵泡颗粒细胞凋亡和自噬相关基因mRNA表达水平的影响

Effects of the FOXO3 Overexpression on the mRNA Expression Level of Apoptosis and Autophagy Related Genes in Goose Follicular Granulosa Cells

本研究首先克隆鹅FOXO3基因编码区(CDS)序列,构建FOXO3基因的真核表达载体,然后将其转染至鹅卵泡颗粒细胞中,利用荧光定量PCR方法检测FOXO3基因对细胞凋亡和自噬相关基因mRNA表达水平的影响。结果显示,本研究成功克隆了鹅FOXO3基因CDS序列,并由此构建了真核表达载体pcDNA3.1-FOXO3。鹅卵泡颗粒细胞FOXO3基因过表达实验发现,过表达组凋亡相关基因Bax、Bcl-2、Caspase3、Fasl的mRNA表达水平较对照组均升高(P<0.05),而自噬相关基因LC3和Beclin1的mRNA表达水平较对照组均降低(P<0.05)。本研究结果表明,FOXO3基因可能影响鹅卵泡颗粒细胞中凋亡及自噬相关基因的表达,进而调控颗粒细胞的凋亡和自噬。

In this study,following the coding region sequence(CDS)of goose FOXO3 gene was cloned,the eukaryotic expression vector was constructed and transfected into goose follicular granulosa cells.Subsequently,the effect of FOXO3 on the mRNA expression level of apoptosis and autophagy related genes was detected by real-time RT-PCR.The results showed that the CDS of goose FOXO3 gene was cloned,and the eukaryotic expression vector pcDNA3.1-FOXO3 was successfully constructed.The overexpression experiment of FOXO3 gene in goose granulosa cells showed that the mRNA expression levels of apoptosis-related genes Bax,Bcl-2,Caspase3 and Fasl in the overexpression group were higher than that in the control group(P<0.05),while the mRNA expression levels of autophagy-related genes LC3 and Beclin1 in the overexpression group were lower than that in the control group(P<0.05).Our study suggested that the FOXO3 gene might influence the expression of apoptosis and autophagy related genes in goose follicular granulosa cells,and thereby regulate the apoptosis and autophagy process of granulosa cells.