猪圆环病毒2型ORF9基因对PK-15细胞的影响
Effect of ORF9 gene of porcine circovirus type 2 on PK-15 cells
边孟婷 1梁海英 1曾智勇 1汤德元 1王彬 1叶泥 1柳佳佳 1黄书 1潘向英 1田红利1
作者信息
- 1. 贵州大学动物科学学院,贵州贵阳 550025
- 折叠
摘要
为探究猪圆环病毒2型(PCV2)ORF9基因对猪肾细胞(PK-15)的影响,通过构建ORF9基因真核表达质粒,并转染至PK-15细胞中,采用流式细胞术及qRT-PCR方法检测过表达ORF9基因对PK-15增殖、凋亡、免疫的影响.结果显示,PCV2 ORF9基因过表达可显著增加内质网应激标志基因GRP78的表达水平;增加PK-15细胞S期细胞数,使细胞增殖速率增快;增加PK-15细胞凋亡率,上调凋亡相关基因caspase-3、caspase-8、caspase-9、p53和Bax的表达水平(P＜0.01),下调凋亡相关基因Bcl-2的表达水平;上调免疫相关基因IL-8、IL-10、NF-KB、TNF-α的表达水平(P＜0.01),下调免疫相关基因IL-2、IFN-β、IL-12的表达水平(P＜0.01).结果表明,PCV2 ORF9基因可能促进PK-15细胞增殖与凋亡,并在PK-15细胞逃逸过程中发挥一定作用.
Abstract
To investigate the effects of ORF9 gene of porcine circovirus type 2(PCV2)on PK-15,eu-karyotic expression plasmid was constructed and transfected into PK-15 cells,and the effects of overexpression of ORF9 on proliferation,apoptosis and immunization of PK-15 cells were exam-ined by flow cytometry and qRT-PCR.The results showed that ORF9 gene overexpression signifi-cantly up-regulated the expression levels of the ER stress marker gene GRP78,increased the num-ber of S phase cells,accelerated cell cycle progression,increased the apoptosis rate of PK-15 cells,up-regulated the expression levels of apoptosis-related genes caspase-3,caspase-8,caspase-9,p53 and Bax(P＜0.01),down-regulated the expression levels of apoptosis-related genes Bcl-2,up-reg-ulated the expression levels of immune-related genes 1L-8,IL-10,NF-κB and TNF-α(P＜0.01),and down-regulated the expression levels of immune-related genes IL-2,IFN-β and IL-12(P＜0.01).The above results indicate that ORF9 gene may promote the proliferation and apoptosis of PK-15 cells and play a role in the escape process of PK-15 cells.
关键词
猪圆环病毒2型/ORF9基因/增殖/凋亡/免疫Key words
PCV2/ORF9 gene/proliferation/apoptosis/immunization引用本文复制引用
基金项目
贵州省科技支撑计划(黔科合支撑[2021]一般162项目)
出版年
2024
NETL
NSTL
万方数据