文章摘要
周 锋,秦 娣,卢 春.HIV-1 Tat对KSHV感染SK-N-SH细胞的影响[J].南京医科大学学报,2009,29(3):296~302
HIV-1 Tat对KSHV感染SK-N-SH细胞的影响
Preliminary research on the effect of HIV-1 Tat upon KSHV infecting SK-N-SH cells
投稿时间:2008-08-29  
DOI:10.7655
中文关键词: 卡波济肉瘤相关疱疹病毒  HIV-1 Tat  共培养
英文关键词: Kaposi’s sarcoma-associated hepersvirus  HIV-1 Tat  co-culture
基金项目:南京医科大学科技发展基金面上项目(08NMUM009)
作者单位
周 锋 南京医科大学微生物学与免疫学系,江苏 南京 210029 
秦 娣  
卢 春  
摘要点击次数: 2232
全文下载次数: 281
中文摘要:
      目的:初步探索人类免疫缺陷病毒1型(HIV-1)反式激活蛋白(Tat)对卡波济肉瘤相关疱疹病毒(KSHV)感染人神经母细胞瘤细胞系SK-N-SH细胞(SK细胞)的影响?方法:将原发性渗出性淋巴瘤(PEL)细胞系BCBL-1细胞与SK细胞共培养,观察SK细胞的细胞病变效应(CPE)并采用Real-time PCR检测KSHV在SK细胞中的复制?重组质粒Tat+pcDNA3.1(+)和空载体pcDNA3.1(+)分别转染SK细胞,经G418筛选获得抗性克隆,以RT-PCR和Western blot检测Tat基因的表达?通过BCBL-1细胞与SK-Tat/SK-vector细胞共培养,采用Real-time PCR检测SK-Tat/SK-vector细胞中KSHV裂解期基因ORF50和26的mRNA表达?结果:细胞共培养后,SK细胞出现CPE且SK细胞中检测到KSHV的基因?RT-PCR和Western blot均在Tat预期位置检测到特异性条带?Real-time PCR检测显示Tat降低KSHV ORF50和26 mRNA转录水平?结论:初步探索表明,在细胞共培养过程中Tat蛋白抑制KSHV在SK细胞中的复制?
英文摘要:
      Objective:To study the effect of human immunodeficiency virus type 1(HIV-1) transactivative transcription protein(Tat) on Kaposi’s sarcoma-associated herpesvirus(KSHV) infecting SK-N-SH neuroblastoma cells(SK cells). Methods:After co-culturing body cavity-based lymphoma 1[BCBL-1, originated from primary effusion lymphoma(PEL)] cells and SK cells, the cytopathic effect(CPE) of SK cells was observed and the replication of KSHV in SK cells was detected by Real-time PCR. SK cells were transfected with recombinant plasmid Tat+pcDNA3.1(+) and empty vector pcDNA3.1(+), respectively. The stably transfected cell clones were obtained through G418 screening. The expression of Tat in SK cells was detected by reverse transcription polymerase chain reaction(RT-PCR) and Western blot. Real-time PCR were further performed to evaluate the expression of KSHV lytic cycle genes ORF50 and ORF26 mRNA after co-culturing BCBL-1 cells with SK-Tat or SK-vector cells. Results:After SK and BCBL-1 cells co-culture, CPE appeared in SK cells and KSHV ORF50 mRNA was expressed in SK cells. The specific band of Tat appeared as expected both in RT-PCR and Western blot detection. Real-time PCR showed that Tat decreased the expression of KSHV ORF50 and ORF26 mRNA. Conclusion:Preliminary research indicated that Tat might inhibit KSHV lytic cycle replication in SK cells during cell co-culture.
查看全文   查看/发表评论  下载PDF阅读器