C8orf4基因编码蛋白对猪流行性腹泻病毒体外复制的抑制效应  

Inhibition Effect of C8orf 4 Gene Encoding Protein on in vitro Replication ofPorcine Epidemic Diarrhea Virus

作　　者：徐红 商红旗[2] 张雪 钱嘉莉[2] 王传红 宋旭 宝梅英 刘诗雨 张格格 郭容利 赵永祥[2] 范宝超 李彬[1,2] XU Hong;SHANG Hongqi;ZHANG Xue;QIAN Jiali;WANG Chuanhong;SONG Xu;BAO Meiying;LIU Shiyu;ZHANG Gege;GUO Rongli;ZHAO Yongxiang;FAN Baochao;LI Bin(College of Veterinary Medicine,Hebei Agricultural University,Baoding 071000,China;Institute of Veterinary Medicine,Jiangsu Academy of Agricultural Sciences,Nanjing 210014,China)

机构地区：[1]河北农业大学动物医学院,保定071000 [2]江苏省农业科学院兽医研究所,南京210014

出　　处：《畜牧兽医学报》2024年第5期2100-2108,共9页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家重点研发计划(2021YFD1801104);国家自然科学基金(32272996)。

摘　　要：C8orf4,也称为甲状腺癌1(thyroid carcinoma 1,TC1),最初是从甲状腺癌及其周围正常甲状腺组织克隆而来,在脊椎动物中普遍表达,具有跨物种的序列保守性。研究表明,C8orf4在肿瘤中高表达,参与各种癌细胞间信息通讯,是一种与癌症和炎症有关的新型调节因子,并通过调节NF-κB的转录增强炎症反应,但对病毒的影响知之甚少。为研究C8orf4编码蛋白对猪流行性腹泻病毒(porcine epidemic diarrhea virus, PEDV)复制的影响,将PEDV接种于Vero细胞,通过qRT-PCR和Western blot检测不同时间点PEDV对C8orf4表达的影响;设计并合成表达C8orf4基因的真核表达载体pcDNA3.1-C8orf44-His和特异性siRNA,通过过表达和抑制C8orf4表达,利用qRT-PCR和Western blot检测C8orf4对PEDV复制的影响;进一步,通过过表达C8orf4编码蛋白,明确C8orf4编码蛋白对PEDV复制周期各阶段的增殖变化。随着PEDV感染时间的增长,细胞中C8orf4表达呈上升趋势;过表达C8orf4显著抑制PEDV复制,而干扰C8orf4表达促进PEDV复制,并且C8orf4编码蛋白主要作用于PEDV复制周期的生物合成阶段。本研究表明C8orf4编码蛋白具有抑制PEDV复制的作用,为C8orf4的功能研究提供参考,为抗PEDV复制机制研究提供基础。C8orf 4,also known as thyroid carcinoma 1(TC1),was originally cloned from thyroid carcinoma and its surrounding normal thyroid tissue.C8orf 4 is universally expressed in vertebrates with sequence conservation across species.Previous studies have shown that C8orf 4 is highly expressed in tumors and is involved in various cancer cell communication.It is a novel regulator of cancer and inflammation,and enhances inflammation by regulating NF-κB transcription.However,little is known about the effects of C8orf 4 on viruses.This experiment was designed to study the effect of C8orf 4 encoding protein on the replication of porcine epidemic diarrhea virus(PEDV).PEDV was inoculated into Vero cells,and the effect of PEDV on C8orf 4 expression was detected by qRT-PCR and Western blot at different time points.The eukaryotic expression vector pcDNA3.1-C8orf 4-His and specific siRNA of C8orf 4 gene were designed and synthesized.The effect of C8orf 4 on PEDV replication was detected by qRT-PCR and Western blot through overexpression and inhibition of C8orf 4 expression.Furthermore,the effect of C8orf 4 encoding protein on the proliferation of PEDV at each stage of the replication cycle was determined by overexpression of C8orf 4 encoding protein.With the extension of virus infection time,the expression of C8orf 4 in cells showed an upward trend.Overexpression of C8orf 4 significantly inhibited PEDV replication,while interfered with C8orf 4 expression to promote PEDV replication,and C8orf 4 encoding protein mainly played a role in the biosynthetic phase of PEDV replication cycle.This study proved that C8orf 4 encoding protein can inhibit PEDV replication,which provides a reference for the study of the function of C8orf 4 and the basis for the study of anti-PEDV replication mechanism.

关 键 词：C8orf4 猪流行性腹泻病毒 抗病毒 复制 

分 类 号：S852.659.6[农业科学—基础兽医学]