表达猫杯状病毒NS2质粒转染猫肾传代细胞的转录组学分析

doi:10.11843/j.issn.0366-6964.2025.12.036

畜牧兽医学报 ›› 2025, Vol. 56 ›› Issue (12): 6339-6350.doi: 10.11843/j.issn.0366-6964.2025.12.036

表达猫杯状病毒NS2质粒转染猫肾传代细胞的转录组学分析

陈尚琛¹^,²(), 董宁宁¹^,²(), 谈晓梅², 李娜², 张琦², 罗茂青², 邓应坤², 庞帅赛², 高嘉溪², 刘光清²^,*(), 孟春春²^,*()

1. 新疆农业大学动物医学学院, 乌鲁木齐 830052
2. 中国农业科学院上海兽医研究所伴侣动物生物安全风险预警与防控技术团队, 上海 200241

收稿日期:2025-02-26 出版日期:2025-12-23 发布日期:2025-12-24
通讯作者: 刘光清,孟春春 E-mail:chenshangchen17@gmail,com;a12367543@outlook.com;liugq@shvri.ac.cn;mengcc@shvri.ac.cn
作者简介:陈尚琛(1999-)，男，江苏盐城人，硕士生，主要从事猫病毒性传染病的致病机制研究，E-mail: chenshangchen17@gmail,com
董宁宁(1996-)，女，河南商丘人，硕士，主要从事猫病毒性传染病的致病机制研究，E-mail: a12367543@outlook.com
第一联系人：
陈尚琛和董宁宁为同等贡献作者
基金资助:
国家自然基金面上项目(32272982)；上海市自然基金项目(23ZR1477100)

Differential Transcriptomic Analysis of Feline Kidney Cells Transfected with Feline Calicivirus NS2 Protein

CHEN Shangchen¹^,²(), DONG Ningning¹^,²(), TAN Xiaomei², LI Na², ZHANG Qi², LUO Maoqing², DENG Yingkun², PANG Shuaisai², GAO Jiaxi², LIU Guangqing²^,*(), MENG Chunchun²^,*()

1. College of Veterinary Medicine, Xinjiang Agricultural University, Urumqi 830052, China
2. Companion Animal Biosafety Risk Warning and Control Technology Team, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, China

Received:2025-02-26 Online:2025-12-23 Published:2025-12-24
Contact: LIU Guangqing, MENG Chunchun E-mail:chenshangchen17@gmail,com;a12367543@outlook.com;liugq@shvri.ac.cn;mengcc@shvri.ac.cn

摘要/Abstract

摘要：

旨在研究FCV NS2蛋白与宿主细胞之间的相互关系。本研究用FCV的NS2真核表达质粒转染CRFK细胞同时设立空载体转染对照组，每组各三个样本于转染后24 h收样进行转录组测序。结果显示：通过比较两组测序结果，发现转染NS2蛋白的试验组细胞存在218个差异表达基因，其中上调基因200个下调基因18个。通过GO富集分析和KEGG富集分析，发现显著差异基因主要集中病毒胞内入侵和免疫逃逸相关信号通路。本研究揭示NS2蛋白对CRFK细胞的内体转运、TGF-β和RIG-Ⅰ等多个信号通路在转录水平具有明显的调控作用，系统性揭示FCV NS2蛋白对宿主细胞整体转录调控模式，为后期研究FCV与宿主细胞互作提供了新的研究思路。

关键词: 猫杯状病毒, NS2, 转录组学测序, 差异表达基因

Abstract:

This study aims to investigate the interaction between FCV NS2 protein and host cells. In this study, a eukaryotic expression plasmid of FCV NS2 was transfected into CRFK cells, with an empty vector as the control. Three samples from each group were collected 24 hours post-transfection for transcriptomic sequencing. Results were as follows: Comparison of the sequencing results revealed 218 differentially expressed genes (DEGs) in the experimental group transfected with NS2 protein, including 200 upregulated genes and 18 downregulated genes. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses showed that the significantly differentially expressed genes were mainly involved in intracellular viral invasion and immune evasion pathways. This study reveals that the NS2 protein significantly regulates multiple signaling pathways at the transcriptional level, including endosomal transport, TGF-β, and RIG-Ⅰ, in CRFK cells, systematically reveals the global transcriptional regulation patterns of host cells by feline calicivirus (FCV) NS2 protein. These findings provide a new perspective for future research on the interactions between FCV and host cells.

Key words: feline calicivirus, NS2, transcriptome sequencing, differentially expressed genes

中图分类号:

S852.659.6

陈尚琛, 董宁宁, 谈晓梅, 李娜, 张琦, 罗茂青, 邓应坤, 庞帅赛, 高嘉溪, 刘光清, 孟春春. 表达猫杯状病毒NS2质粒转染猫肾传代细胞的转录组学分析[J]. 畜牧兽医学报, 2025, 56(12): 6339-6350.

CHEN Shangchen, DONG Ningning, TAN Xiaomei, LI Na, ZHANG Qi, LUO Maoqing, DENG Yingkun, PANG Shuaisai, GAO Jiaxi, LIU Guangqing, MENG Chunchun. Differential Transcriptomic Analysis of Feline Kidney Cells Transfected with Feline Calicivirus NS2 Protein[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(12): 6339-6350.

图/表 10

表 1

表 2

图 1

图 2

图 3

图 4

表 3

表达差异显著的基因信息表"

基因名称 Gene name	基因ID Gene ID	基因描述 Gene description	差异倍数 FC(ns2/myc)	P值 P value	调节 Regulate
SLC7A11	101 099 374	溶质载体家族成员SLC7A11 Solute carrier family 7 member 11	3.087	0.000 000	up
GREM1	101 096 735	Gremlin 1(DAN家族BMP拮抗剂) Gremlin 1, DAN family BMP antagonist	3.012	0.000 000	up
STEAP2	2 130 929 580	STEAP2金属还原酶 STEAP2 metalloreductase	2.878	0.000 003	up
CCPG1	2 130 984 062	细胞周期进程展1 Cell cycle progression 1	2.589	0.000 714	up
RIF1	2 131 005 132	复制时间调节因子1 Replication timing regulatory factor 1	2.550	0.003 134	up
ESCO2	2 130 950 868	姐妹染色单体凝聚N-乙酰转移酶2的建立 Establishment of sister chromatid cohesion N-acetyltransferase 2	2.485	0.000 024	up
BRCA2	57 619 070	BRCA2 DNA修复相关 BRCA2 DNA repair associated	2.478	0.000 111	up
NEMF	101 093 444	核输出介质因子 Nuclear export mediator factor	2.466	0.001 278	up
SYTL2	2 131 026 479	突触结合蛋白样2 Synaptotagmin like 2	2.428	0.000 013	up
EEA1	101 088 764	早期内体抗原1 Early endosome antigen 1	2.416	0.004 651	up
ABCB4	2 130 921 776	ATP结合盒亚家族B成员4 ATP binding cassette subfamily B member 4	0.125	0.000 000	down
EGR1	101 088 388	早期生长反应1 Early growth response 1	0.212	0.000 000	down
USP18	101 093 867	泛素特异性肽酶18 Ubiquitin specific peptidase 18	0.384	0.000 036	down
CD74	101 085 953	CD74分子 CD74 molecule	0.398	0.012 070	down
ESRRB	2 130 992 031	雌激素相关受体β Estrogen related receptor beta	0.400	0.000 000	down
MX1	101 085 996	MX动力样蛋白GTP酶1 MX dynamin like GTPase 1	0.403	0.000 002	down
CHRNB4	2 130 977 942	胆碱能受体烟碱β4亚基 Cholinergic receptor nicotinic beta 4 subunit	0.436	0.000 000	down
EPHB2	101 095 030	酪氨酸蛋白激酶受体B2 EPH receptor B2	0.460	0.000 000	down
IRF7	101 084 060	干扰素调节因子7 Interferon regulatory factor 7	0.465	0.000 000	down
DHX58	101 094 463	DExH盒解旋酶58 DExH-box helicase 58	0.466	0.000 236	down

表 3

图 5

图 6

图 7

参考文献 24

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引物名称 Primer name	引物序列(5′→3′) Primer sequence
CMV-NS2-F	gaattcggtcgaccgagatctctATGGCTTGTCCATCATGTGCT
CMV-NS2-R	catgtctggatccccgcggccgcCTCTGATCTAAATCCAGTGAATAGAGCT

引物名称 Primer name	引物序列(5′→3′) Primer sequence
q-NEMF-F	AGGTGGAAGAGATCAGCAACAG
q-NEMF-R	GGAATGGGTTCTCCTGTTGGAT
q-SYTL2-F	TGTCCCAGGAAAACACAAGGAA
q-SYTL2-R	TGATCTTCTGGGAACTTGGAGC
q-EEA1-F	ACTGAGAAGGAGGCTAAACTGC
q-EEA1-R	AGACTGCCACTCCTTCTTTACG
q-ESCO2-F	ACCCGATCTTCAGTGTGTCTTC
q-ESCO2-R	AATCTGTGGTGATGCTGGACAT
q-CCPG1-F	GAATCTTCCACAGCTTCTCCCA
q-CCPG1-R	AAGTCGGCACTTGAGAGAGAAG
q-SLC7A11-F	CTAACCAAGAAAGGGGGCATCT
q-SLC7A11-R	AAAGGGGCATGTCTAAGTGAGG
q-GREM1-F	TTCAGTCCTGCTCTTTCTGCAA
q-GREM1-R	TATGCATCGACACTGCTTGACT
q-RIF1-F	AGAAACCAGCCCAGACATACTG
q-RIF1-R	GTGGGGGAGTTCCAGAAATGAT
q-BRCA2-F	GCACTACAAAAGGCAAGACAGG
q-BRCA2-R	TCCCACTTGCTGTACTGAATCC
q-STEAP2-F	ACCTTGCCCATAGTTGCCATTA
q-STEAP2-R	TTCCTCGTTCCAAGAGTTCTCG
q-USP18-F	GGGAACAGAAGAGCAAAGGAGA
q-USP18-R	GGAGACAGTAGGCAAGTTCCAA
q-MX1-F	TGATATGGTCCAGCTTGCCTTT
q-MX1-R	ATTCCTCTTCTCCTCCTCCGAA
q-EPHB2-F	ACAAGATGATCCGAAACCCCAA
q-EPHB2-R	ACTCATCCACGGTGTTAAAGCT
q-ABCB4-F	GGTGTTTCAGAATGGCAAGGTC
q-ABCB4-R	CATAAGTTCTGTGTCCCAGCCT
q-EGR1-F	CCACCACCTCTCTCTCTTCCTA
q-EGR1-R	TGAGGAACCAGGAGAGGAGTAG
q-DHX58-F	AACCCCAAACTGGAGATGCTAG
q-DHX58-R	TCTCTCTGGGTCATATGGGTGT
q-CD74-F	CCTTTGAACAAGTTGCGGGTAG
q-CD74-R	CTCCAGGGAGTTCTTGCTCATT
q-CHRNB4-F	TATTGCCCAGCACATGAAGAGT
q-CHRNB4-R	GAGCACACAGACAACCACAAAG
q-ESRRB-F	AAGAAGCTCAAGGTGGAGAAGG
q-ESRRB-R	CCAGCATCTCCAGGAAGAGTTT
q-IRF7-F	AGCTTCGTGTTTCTGTATGGGT
q-IRF7-R	CCTCCCAGTAGACCTTGCATTT

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表达猫杯状病毒NS2质粒转染猫肾传代细胞的转录组学分析

Differential Transcriptomic Analysis of Feline Kidney Cells Transfected with Feline Calicivirus NS2 Protein

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