Please wait a minute...

中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志
中国生物工程杂志  2021, Vol. 41 Issue (2/3): 7-13    DOI: 10.13523/j.cb.2009002
研究报告     
人呼吸道合胞病毒感染的A549细胞中长链非编码RNA表达谱研究
刘美琴,高博,焦月盈,李玮,虞结梅,彭向雷,郑妍鹏,付远辉,何金生()
北京交通大学生命科学与生物工程研究院 北京 100044
Long Non-coding RNA Expression Profile in A549 Cells Infected with Human Respiratory Syncytial Virus
LIU Mei-qin,GAO Bo,JIAO Yue-ying,LI Wei,YU Jie-mei,PENG Xiang-lei,ZHENG Yan-peng,FU Yuan-hui,HE Jin-sheng()
College of Life Sciences & Bioengineering, Beijing Jiaotong University, Beijing 100044, China
 全文: PDF(23754 KB)   HTML
摘要：

目的: 探讨人呼吸道合胞体病毒(human respiratory syncytial virus,RSV)感染A549细胞的长链非编码RNA(long non-coding RNAs, lncRNA)表达谱的差异,更好地了解宿主与RSV之间的相互作用的可能分子机制。方法: 1 MOI RSV感染A549细胞,24h后提取RNA,利用Agilent的lncRNA表达谱芯片,筛查了RSV感染和未感染的A549细胞中的lncRNA差异表达谱,并进行实时定量PCR验证,通过GO和KEGG对差异mRNA和lncRNA进行生物信息学分析。 结果: RSV感染的A549细胞与未感染的A549细胞相比,共有1 463个lncRNA和1 552个mRNAs显著上调,有944个lncRNA和1 489个mRNAs下调,差异表达lncRNA及其预测靶基因主要富集于免疫应答过程。通过定量PCR证实所选lncRNA的表达谱结果。结论: RSV感染A549细胞后,lncRNA表达谱发生明显变化,对深入研究lncRNA如何参与RSV与宿主之间的相互作用奠定了基础。
关键词： 人呼吸道合胞体病毒;  A549细胞;  长链非编码RNA;  表达谱    
Abstract:

Objective: To better understand the possible molecular mechanism of the interaction between the host and human respiratory syncytial virus (RSV), the long non-coding RNA expression profile in A549 cells infected by RSV was investigated. Methods: After 24 hours of A549 cells infected with 1 MOI RSV and extracted for RNA samples, lncRNA differential expression profile was screened by Agilent’s lncRNA expression profile chip and compared with the A549 cell control. LncRNA differential expression profile was verified by real-time quantitative PCR (qRT-PCR). Bioinformatics analysis of differential mRNA and lncRNA was performed by GO and KEGG. Results: A total of 1 463 lncRNA and 1 552 mRNAs were significantly up-regulated in RSV-infected A549 cells compared with A549 cell control, while 944 lncRNA and 1 489 mRNAs were down-regulated. LncRNA and the predicted target genes were mainly enriched in the immune response. The expression profile of the selected lncRNA was confirmed by qRT-PCR. Conclusion: LncRNA expression profile changes significantly after RSV infection of A549 cells, but the underlining mechanism in the interaction between RSV and host needs to be further explored.
Key words: Human respiratory syncytial virus    A549 cells    Long non-coding RNA    Expression profile
收稿日期: 2020-09-01 出版日期: 2021-04-08
ZTFLH:  Q812  
通讯作者: 何金生     E-mail: jshhe@bjtu.edu.cn
服务　　
把本文推荐给朋友
加入引用管理器
E-mail Alert
RSS
作者相关文章　　
刘美琴
高博
焦月盈
李玮
虞结梅
彭向雷
郑妍鹏
付远辉
何金生

引用本文:

刘美琴,高博,焦月盈,李玮,虞结梅,彭向雷,郑妍鹏,付远辉,何金生. 人呼吸道合胞病毒感染的A549细胞中长链非编码RNA表达谱研究[J]. 中国生物工程杂志, 2021, 41(2/3): 7-13.

LIU Mei-qin,GAO Bo,JIAO Yue-ying,LI Wei,YU Jie-mei,PENG Xiang-lei,ZHENG Yan-peng,FU Yuan-hui,HE Jin-sheng. Long Non-coding RNA Expression Profile in A549 Cells Infected with Human Respiratory Syncytial Virus. China Biotechnology, 2021, 41(2/3): 7-13.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.2009002    或    https://manu60.magtech.com.cn/biotech/CN/Y2021/V41/I2/3/7

RSV感染的A549细胞中上调的lncRNAs RSV感染的A549细胞中下调的lncRNAs
lncRNA ID FC P value lncRNA ID FC P value
ENST000007613.1 168.389 0.000 594 ENST00000520156.1 22.824 0.000 992
RNA147145|p0249_imsncRNA392 117.749 0.000 083 ENST00000579368.1 13.528 0.000 106
ENST00000415810.1 92.435 0.000 213 uc010lcl.3 11.711 0.000 133
HIT000071394 85.878 0.000 213 ENST00000560973.1 10.848 0.006 189
uc003ylb.1 82.100 0.000 424 ENST00000560237.1 9.344 0.000 017
TCONS_00027056 71.146 0.000 215 ENST00000599209.1 8.865 0.000 839
XR_426856.1 70.406 0.000 122 XR_429553.1 8.307 0.000 789
ENST00000600152.1 66.528 0.000 038 TCONS_00028233 8.162 0.005 519
ENST00000592728.1 65.052 0.000 355 ENST00000521373.1 7.929 0.003 902
ENST00000595705.1 59.927 0.001 780 XR_247160.1 7.482 0.000 099
表1  与未感染的A549细胞相比,RSV感染的A549细胞中10个上调和下调最显著的lncRNAs
RSV感染的A549细胞中上调的mRNAs RSV感染的A549细胞中下调的mRNAs
Genbank Accession FC P value Genbank Accession FC P value
NM_172139 4 903.417 0.000 011 NM_001005190 34.342 0.000 026
NM_001001437 1 914.344 0.000 012 NM_014405 14.089 0.007 145
NM_172140 1 675.346 0.000 076 NM_207362 13.127 0.000 356
NM_005409 1 025.258 0.000 089 NM_001001925 11.840 0.000 955
NM_017523 469.671 0.000 023 NM_001145064 10.694 0.000 069
NM_001565 457.013 0.000 045 NM_004975 10.657 0.000 007
NM_001565 374.110 0.000 019 NM_021871 10.468 0.002 490
NM_080657 330.406 0.000 049 NM_001736 10.147 0.006 427
NM_017523 288.632 0.000 007 NM_032294 9.232 0.000 058
XM_006724857 278.801 0.000 170 NM_014376 9.077 0.000 063
表2  与未感染的A549细胞相比,RSV感染的A549细胞中10个上调和下调最显著的mRNAs
图1  差异表达的lncRNA和mRNA聚类分析
图2  IncRNA调控的mRNA的GO和pathway富集分析结果
图3  18种显著差异表达的lncRNAs的qRT-PCR验证结果
图4  5种显著差异表达的mRNAs的qRT-PCR验证结果
图5  敲低lncRNA对A549细胞蛋白质表达水平的影响
[1] Thornhill E M, Verhoeven D. Respiratory syncytial virus’s non-structural proteins: masters of interference. Frontiers in Cellular and Infection Microbiology, 2020,10:225.
doi: 10.3389/fcimb.2020.00225 pmid: 32509597
[2] Hu M J, Bogoyevitch M A, Jans D A. Impact of respiratory syncytial virus infection on host functions: implications for antiviral strategies. Physiological Reviews, 2020,100(4):1527-1594.
doi: 10.1152/physrev.00030.2019 pmid: 32216549
[3] Zhou Y, Li M, Xue Y, et al. Interferon-inducible cytoplasmic lncLrrc55-AS promotes antiviral innate responses by strengthening IRF3 phosphorylation. Cell Research,, 2019,29(8):641-654.
doi: 10.1038/s41422-019-0193-0 pmid: 31213650
[4] Swedan S, Musiyenko A, Barik S. Respiratory syncytial virus nonstructural proteins decrease levels of multiple members of the cellular interferon pathways. Journal of Virology, 2009,83(19):9682-9693.
doi: 10.1128/JVI.00715-09 pmid: 19625398
[5] Kulkarni S, Lied A, Kulkarni V, et al. CCR5AS lncRNA variation differentially regulates CCR5, influencing HIV disease outcome. Nature Immunology, 2019,20(7):824-834.
doi: 10.1038/s41590-019-0406-1 pmid: 31209403
[6] Ahmed W, Liu Z F. Long non-coding RNAs: novel players in regulation of immune response upon herpesvirus infection. Frontiers in Immunology, 2018,9:761.
doi: 10.3389/fimmu.2018.00761 pmid: 29706968
[7] Kim J C, Ha Y J, Tak K H, et al. Opposite functions of GSN and OAS2 on colorectal cancer metastasis, mediating perineural and lymphovascular invasion, respectively. PLoS One, 2018,13(8):e0202856.
doi: 10.1371/journal.pone.0202856 pmid: 30148861
[8] Wu S Q, Wang Y, Chen G, et al. 2',5'-Oligoadenylate synthetase 1 polymorphisms are associated with tuberculosis: a case-control study. BMC Pulmonary Medicine, 2018,18(1):1-10.
doi: 10.1186/s12890-017-0557-5 pmid: 29301525
[9] Jin S H, Cui J. BST2 inhibits type I IFN (interferon) signaling by accelerating MAVS degradation through CALCOCO2-directed autophagy. Autophagy, 2018,14(1):171-172.
doi: 10.1080/15548627.2017.1393590 pmid: 29165031
[10] Jin S H, Tian S, Luo M, et al. Tetherin suppresses type I interferon signaling by targeting MAVS for NDP52-mediated selective autophagic degradation in human cells. Molecular Cell, 2017,68(2):308-322 e4.
doi: 10.1016/j.molcel.2017.09.005 pmid: 28965816
[1] 刘天义,冯卉,SALSABEELYousuf,解领丽,苗向阳. lncRNA在动物脂肪沉积中的研究进展*[J]. 中国生物工程杂志, 2021, 41(11): 82-88.
[2] 万群,刘梦瑶,夏菁,苟理尧,唐敏,孙恃雷,张彦. 长链非编码RNA SNHG3对人乳腺癌细胞MCF-7增殖、迁移与侵袭的影响 *[J]. 中国生物工程杂志, 2019, 39(1): 13-20.
[3] 谭杨,刘胜,罗凤玲,章晓联. 结核分枝杆菌H37Rv刺激巨噬细胞后差异表达lncRNA分析及鉴定 *[J]. 中国生物工程杂志, 2018, 38(5): 1-9.
[4] 范源,罗嘉,甘麦邻,谭娅,张顺华,朱砺. 长链非编码RNA TERRA的研究进展 *[J]. 中国生物工程杂志, 2018, 38(10): 64-73.
[5] 辛娟, 林福春, 张兵兵, 向燕, 王远亮. 力生长因子E肽对成骨细胞增殖及基因表达的影响[J]. 中国生物工程杂志, 2011, 31(10): 1-6.
[6] 万丙良 査中萍 杜雪树. 水稻热激转录因子基因在植物激素和非生物环境胁迫条件下的表达谱分析[J]. 中国生物工程杂志, 2010, 30(10): 0-0.
[7] 万丙良, 査中萍, 杜雪树. 水稻热激转录因子基因在植物激素和非生物环境胁迫条件下的表达谱分析[J]. 中国生物工程杂志, 2010, 30(10): 22-32.
[8] 尤元海 张建中. 基因表达谱芯片的数据挖掘[J]. 中国生物工程杂志, 2009, 29(10): 87-91.
[9] 王晚霞 兰喜 徐向红 居军 柳纪省. 颗粒溶素和穿孔素真核共表达及其对A549细胞的影响[J]. 中国生物工程杂志, 2009, 29(10): 12-17.
[10] 杨静,顾玉超,于文功. Twist对小鼠乳腺癌细胞基因表达谱的调控研究[J]. 中国生物工程杂志, 2008, 28(4): 1-6.
[11] 狄春红, 杨磊, 和桂芬, 仙玲, 谭晓华, 吴奇涵, 季朝能, 顾少华. 凋亡和周期基因芯片研究As2O3对NB4细胞凋亡相关基因表达谱的影响[J]. 中国生物工程杂志, 2005, 25(11): 12-16.
[12] 饶志明, 张荣珍, 王正祥, 方慧英, 诸葛健. 基因芯片技术在微生物学研究中的应用[J]. 中国生物工程杂志, 2003, 23(8): 61-65.
[13] 王升启. 基因芯片技术及应用研究进展[J]. 中国生物工程杂志, 1999, 19(4): 45-51.
主管：中国科学院 　主办：中国科学院文献情报中心 　中国生物技术发展中心 　中国生物工程学会