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高表达CD81的Huh7细胞株对提高HCV病毒感染效率的研究 被引量:1
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作者 杨硕 徐珩 +7 位作者 顾娜 冯丹丹 李宁 李军锋 童贻刚 周育森 娄金丽 闾军 《北京医学》 CAS 2010年第6期424-427,共4页
目的建立具有高感染性的丙型肝炎病毒(HCV)的体外细胞株。方法构建高表达CD81的Huh7细胞株。将HCV质粒(JFH-1,2a)体外转录成RNA,电转染到Huh7和CD81-Huh7细胞中,实时定量PCR法检测两组细胞和培养上清液中病毒载量,间接免疫荧光法检测细... 目的建立具有高感染性的丙型肝炎病毒(HCV)的体外细胞株。方法构建高表达CD81的Huh7细胞株。将HCV质粒(JFH-1,2a)体外转录成RNA,电转染到Huh7和CD81-Huh7细胞中,实时定量PCR法检测两组细胞和培养上清液中病毒载量,间接免疫荧光法检测细胞中HCV核心蛋白表达。收集电转后细胞上清液重新感染两组细胞,检测细胞中病毒RNA表达。结果 CD81-Huh7电转后,细胞和上清液中病毒RNA水平均明显高于Huh71-2log。其被感染的效率也高于Huh7。结论成功建立一株能够高表达感染性HCV病毒颗粒的细胞株CD81-Huh7。 展开更多
关键词 丙型肝炎病毒 CD81 JFH-1 huh7细胞
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Different Responses of Two Highly Permissive Cell Lines Upon HCV Infection 被引量:2
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作者 Honghe Chen Rongjuan Pei Xinwen Chen 《Virologica Sinica》 SCIE CAS CSCD 2013年第4期202-208,共7页
The construction of the first infectious clone JFH-1 speeds up the research on hepatitis C virus (HCV). However, Huh7 cell line was the only highly permissive cell line for HCV infection and only a few clones were ful... The construction of the first infectious clone JFH-1 speeds up the research on hepatitis C virus (HCV). However, Huh7 cell line was the only highly permissive cell line for HCV infection and only a few clones were fully permissive. In this study, two different fully permissive clones of Huh7 cells, Huh7.5.1 and Huh7-Lunet-CD81 (Lunet-CD81) cells were compared for their responses upon HCV infection. The virus replication level was found slightly higher in Huh7.5.1 cells than that in Lunet-CD81 cells. Viability of Huh7.5.1 cells but not of Lunet-CD81 cells was reduced significantly after HCV infection. Further analysis showed that the cell cycle of infected Huh7.5.1 cells was arrested at G1 phase. The G1/S transition was blocked by HCV infection in Huh7.5.1 cells as shown by the cell cycle synchronization analysis. Genes related to cell cycle regulation was modified by HCV infection and gene interaction analysis in GeneSpring GX in Direct Interactions mode highlighted 31 genes. In conclusion, the responses of those two cell lines were different upon HCV infection. HCV infection blocked G1/S transition and cell cycle progress, thus reduced the cell viability in Huh7.5.1 cells but not in Lunet-CD81 cells. Lunet-CD81 cells might be suitable for long term infection studies of HCV. 展开更多
关键词 HCV huh7.5.1 huh7-lunet-cd81 VIABILITY Cell cycle G1/S
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