| 宿主因子CD63对人轮状病毒SA11体外复制的影响 |
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| 引用本文： | 郑春红,于漫,李香雨,聂梓陶,赵微.宿主因子CD63对人轮状病毒SA11体外复制的影响[J].现代预防医学,2020,0(12):2233-2236. |
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| 作者姓名： | 郑春红 于漫 李香雨 聂梓陶 赵微 |
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| 作者单位： | 锦州医科大学，辽宁 锦州 121001 |
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| 摘 要： | 目的 测定轮状病毒2种宿主细胞内CD63的高低表达对病毒体外复制的影响，为探究轮状病毒与宿主因子CD63的相互作用机制奠定基础。方法 构建CD63沉默表达质粒shRNA - 4038、shRNA - 4041和过表达质粒pcDNA3.1（+） - CD63 - flag（CD63 - Flag）。质粒转染轮状病毒2种宿主细胞CV - 1和MA104，采用实时定量PCR（RT - qPCR）和western blot检测CD63 mRNA和蛋白的表达，筛选CD63高表达细胞株（CD63 - H）和低表达细胞株(CD63 - L)。轮状病毒SA11感染CD63 - L、CD63 - H、正常对照细胞组（CNr）及空白对照细胞组（shNC），采用RT - qPCR、western blot检测病毒基因拷贝数和病毒蛋白表达。结果 细胞转染shRNA - 4038和shRNA - 4041质粒后， CD63的表达明显抑制，其中CV - 1细胞mRNA和蛋白水平抑制效率分别为69.7%和39.8%，MA104细胞mRNA和蛋白水平抑制效率分别为76.4%和49.3% ；CD63 - H组细胞内CD63的mRNA和蛋白表达明显增加（P＜0.05）。采用滴度为8logFFU/ml的SA11病毒感染细胞后，CD63 - L组病毒基因拷贝数下降32.6%（CV - 1）和35.7%（MA104）；病毒蛋白表达水平下降41.6%（CV - 1）和44.7%（MA104）；CD63 - H组病毒基因拷贝数和蛋白表达水平明显增加，对照组病毒表达量无明显变化（P＜0.05）。结论 宿主因子CD63高表达能够促进病毒复制，而CD63抑制表达后，病毒复制明显下降。宿主因子CD63的表达与轮状病毒复制呈现正相关。
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| 关 键 词： | CD63蛋白 轮状病毒 病毒复制 |
Effect of host factor CD63 on replication of human rotavirus SA11 in vitro |
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| ZHENG Chun-hong,YU Man,LI Xiang-yu,NIE Zi-tao,ZHAO Wei.Effect of host factor CD63 on replication of human rotavirus SA11 in vitro[J].Modern Preventive Medicine,2020,0(12):2233-2236. |
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| Authors: | ZHENG Chun-hong YU Man LI Xiang-yu NIE Zi-tao ZHAO Wei |
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| Affiliation: | Jinzhou Medical University, Jinzhou, Liaoning 121001, China |
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| Abstract: | To determine the effect of high and low expression of CD63 in two host cells of rotavirus on virus replication in vitro, and to lay a foundation for exploring the interaction mechanism of rotavirus and host factor CD63.Methods CD63 silent expression plasmids shRNA-4038, shRNA-4041 and overexpression plasmid pcDNA3.1(+)-CD63-flag(CD63-Flag) were constructed. The plasmid was transfected into two host cells of rotavirus, CV-1 and MA104, and the expression of mRNA and protein of CD63 was detected by real-time quantitative PCR(RT-qPCR) and western blot. CD63 over-expressing cell lines(CD63-H) and low expression cell lines(CD63-L) were screened. Rotavirus SA11 infected CD63-L, CD63-H, normal control cell group(CNr) and blank control cell group(shNC), RT-qPCR and western blot were used to detect virus gene copy number and viral protein expression. Results After the cells were transfected with shRNA-4038 and shRNA-4041 plasmids, the expression of CD63 was significantly inhibited. Among them, the inhibition efficiency of mRNA and protein levels of CV-1 cells were 69.7% and 39.8%, and the inhibition efficiency of mRNA and protein levels of MA104 cells were 76.4% and 49.3%, respectively; the mRNA and protein expression of CD63 in CD63-H cells were significantly increased(P<0.05). After infecting cells with SA11 virus with a titer of 8 logFFU/ml, the copy number of virus genes in the CD63-L group decreased by 32.6%(CV-1) and 35.7%(MA104); viral protein expression levels decreased by 41.6%(CV-1)and 44.7%(MA104), respectively. CD63-H group virus gene copy number and protein expression level increased significantly, the control group virus expression level did not change significantly(P<0.05). Conclusion The high expression of host factor CD63 can promote virus replication, while inhibition of expression of host factor CD63 can significantly reduce virus replication. The expression of host factor CD63 is positively correlated with rotavirus replication. |
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| Keywords: | CD63 protein Rotavirus Virus replication |
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