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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OA北大核心CSTPCD

Inhibition Effect of C8orf4 Gene Encoding Protein on in vitro Replication of Porcine Epidemic Diarrhea Virus

中文摘要英文摘要

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.

C8orf4,also known as thyroid carcinoma 1(TC1),was originally cloned from thyroid carcinoma and its surrounding normal thyroid tissue.C8orf4 is universally expressed in vertebrates with sequence conservation across species.Previous studies have shown that C8orf4 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 C8orf4 on viruses.This experiment was designed to study the effect of C8orf4 encoding protein on the replication of porcine epidemic diarrhea virus(PEDV).PEDV was inoculated into Vero cells,and the effect of PEDV on C8orf4 expression was detected by qRT-PCR and Western blot at different time points.The eukaryotic expression vector pcDNA3.1-C8orf4-His and specific siRNA of C8orf4 gene were designed and synthesized.The effect of C8orf4 on PEDV replication was detected by qRT-PCR and Western blot through overexpression and inhibition of C8orf4 expression.Furthermore,the effect of C8orf4 encoding protein on the proliferation of PEDV at each stage of the replication cycle was determined by overexpression of C8orf4 encoding protein.With the extension of virus infection time,the expression of C8orf4 in cells showed an upward trend.Overexpression of C8orf4 significantly inhibited PEDV replication,while interfered with C8orf4 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 C8orf4 and the basis for the study of anti-PEDV replication mechanism.

徐红;郭容利;赵永祥;范宝超;李彬;商红旗;张雪;钱嘉莉;王传红;宋旭;宝梅英;刘诗雨;张格格

河北农业大学动物医学院,保定 071000||江苏省农业科学院兽医研究所,南京 210014江苏省农业科学院兽医研究所,南京 210014

畜牧业

C8orf4猪流行性腹泻病毒抗病毒复制

C8orf4PEDVanti-virusreplication

《畜牧兽医学报》 2024 (005)

2100-2108 / 9

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

10.11843/j.issn.0366-6964.2024.05.028

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