巴马小型猪内源性反转录病毒的整合对HEK293细胞周期、细胞凋亡和相关基因表达的影响  

Effects of the integraton of PERV from inbreeded Bama miniature pigs on cell cycle and apoptosis as well as the expresson of some related genes in HEK293 cells

作　　者：马玲 钟华 秦树英 唐海波 白安斌 陈凤莲 刘金凤 覃绍敏 吴健敏 MA Ling;ZHONG Hua;QIN Shu-ying;TANG Hai-bo;BAI An-bin;CHEN Feng-lian;LIU Jin-feng;QIN Shao-min;WU Jian-min(College of Animal Science and Technology,Guangxi University,Nanning 530004,China;Guangxi Key Laboratory of Veterinary Biotechnology,Guangxi Veterinary Research Institute,Nanning 530001,China;College of Ocean and Biotechnology,Guangxi University for Nationalities,Nanning 530006,China)

机构地区：[1]广西大学动物科学技术学院,广西南宁530004 [2]广西壮族自治区兽医研究所,广西兽医生物技术重点实验室,广西南宁530001 [3]广西民族大学海洋与生物技术学院,广西南宁530006

出　　处：《中国兽医学报》2020年第5期882-887,共6页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31260613);广西自然科学基金资助项目(2015GXNSFAA139089,2013GXNSFBA019123);广西兽医生物技术重点实验室开放基金资助项目(14-045-31-B-5,12-071-28-B-3)。

摘　　要：检测广西巴马小型猪内源性反转录病毒(PERV)整合到HEK293细胞基因组(HEK293-PERV-BM)后,宿主细胞的细胞周期、细胞凋亡及cyclin D1、CDK4、c-Myc、p53、p16和k-Ras等相关基因表达的改变,推测相关作用机制。检测发现P10代巴马小型猪内源性反转录病毒感染HEK293细胞模型(HEK293-PERV-BM)的细胞周期主要被阻滞在S期和G2/M期,细胞凋亡受到抑制,P25代细胞周期主要被阻滞在S期,细胞凋亡明显受到诱导。通过实时荧光定量PCR和Western blot检测发现细胞周期相关基因的表达与细胞感染模型传代次数(P1~P35)有密切关系,其中cyclin D1、c-Myc和k-Ras基因表现为先降低后升高,而CDK4和p16基因则恰好相反,同时,p53基因表达的改变不明显,据此可以推测P10代细胞周期的改变可能由Rb调控通路诱导发生,P25代细胞周期的改变可能由Rb调控通路与p53调控通路协同诱导发生。上述结果提示PERV的感染可能存在潜在的危险。The cell cycle and apoptosis as well as the expression of some related genes were altered after the integration of PERV-BM,and differed with different passages of HEK293-PERV-BM cells(P1-P35).In HEK293-PERV-BM P10,the cell cycle was arrested in S phase and G2/M phase as well as the cell apoptosis was inhibited.In HEK293-PERV-BM P25,the cell cycle was arrested in S phase and the apoptosis was induced.To find the mechanisms,qPCR was applied to detect the mRNA expression of cyclin D1,CDK4,c-Myc,p53,p16 and k-Ras genes in different passages of HEK293-PERV-BM.Meanwhile,the protein expressions of cyclin D1,CDK4,c-Myc and other genes was detected by Western blot.The expression level of cyclin D1,c-Myc and k-Ras genes exhibited decrease and then increase,while the expression of CDK4 and p16 genes did just the opposite,and p53 did not significantly change.The expressions of the related genes suggested that the cell cycle of lower passages might be regulated by Rb passway,while the cell cycle of higher passages might be regulated by Rb passway and p53 passway.These results provide the clues of potential infection and effects of PERV-BM.

关 键 词：巴马小型猪 猪内源性反转录病毒 细胞周期 细胞凋亡 

分 类 号：S852.65[农业科学—基础兽医学]