目的检测EGFR基因在新疆卡波西肉瘤（Kaposi’s Sarcoma，KS）中的表达并分析其与KS发生发展的关系。方法应用SYBR Green I作为荧光染料，以GAPDH作内参照，建立检测EGFR mRNA表达的实时荧光PCR反应体系，对经病理活检证实的7例KS肿瘤...目的检测EGFR基因在新疆卡波西肉瘤（Kaposi’s Sarcoma，KS）中的表达并分析其与KS发生发展的关系。方法应用SYBR Green I作为荧光染料，以GAPDH作内参照，建立检测EGFR mRNA表达的实时荧光PCR反应体系，对经病理活检证实的7例KS肿瘤组织和瘤旁正常组织中EGFR mRNA的表达水平进行检测。结果KS肿瘤组织中EGFR表达水平低于瘤旁正常组织（t=-3．865，P〈0．05），并且结果有统计学意义。KS肿瘤组织中EGFR的相对表达量为0．34±0．31，瘤旁组织中为1．24±0．78（P=0．008）。结论所建立的SYBR Green Ⅰ实时定量PCR方法可以成功地检测EGFR基因的表达量。KS肿瘤组织中EGFR表达水平低于瘤旁正常组织，差异有统计学意义。展开更多
Objective: To study and confirm that recombinant cytokines similar to those produced by HIV-1 infected T cells induced lytic cycle replication of human herpesvirus 8 (HHV-8) in BC-3 cells, another cell line from prima...Objective: To study and confirm that recombinant cytokines similar to those produced by HIV-1 infected T cells induced lytic cycle replication of human herpesvirus 8 (HHV-8) in BC-3 cells, another cell line from primary effusion lymphama(PEL). Methods: The persistent stimulation of BC-3 was conducted by several cytokines known to be produced by HIV-1-infected T cells and important in growth and proliferation of Kaposi's sarcoma(KS)cells in vitro, such as the interferon-γ (IFN-γ) , tlie hepatocyte growth factor/scatter factor (HGF / SF) , the Oncostain M(OSM) , and the tumor necrosis factor-α (TNF-α)which is not produced by HIV-1-infected T cells. Treated and untreated BC-3 cells were collected at the 3rd and 7th day of persistent stimulation, respectively. Immuno-histochemical (IHC) staining, Northern blot, quantitative PCR (real- time PCR ) and electron microscopy (EM) were carried out to detect the expression of immunogenic protein ORF59, messenger RNA (mRNA) of minor capsid protein ORF26, and the presence of viral particles of HHV-8 from treated and untreated BC-3 cells. Results: It showed that IFN-γ, HGF/SF, OSM, and TNF-α were found to induce an increase in mRNA expression of ORF26 when added individually to BC-3 cells. Particularly, ORF26 expression stimulated with IFN-γ and TNF-α respectively, increased 6. 1 and 2. 5-fold(from real-time PCR results)at the 7th day when compared with untreated BC-3 cells. Meanwhile, about 20% of IFN-γ stimulated BC-3 cells expressed ORF59 at the 7th day as compared with 1. 5% of untreated BC-3 cells when IHC staining was employed. In addition, viral particles of HHV-8 were readily identified in BC-3 cells stimulated with IFN-γ at the 7th day with EM analysis. Conclusion;TNF-α and recombinant cytokines being similar to those produced by HIV- 1 infected T Cells could really induce HHV- 8 lytic cycle replication in BC-3 cells, another cell line of PEL.展开更多
Kaposi’s sarcoma-associated herpesvirus(KSHV), also known as human herpesvirus-8(HHV-8), is etiologically linked to the development of Kaposi’s sarcoma, primary effusion lymphoma, and multicentric Castleman’s disea...Kaposi’s sarcoma-associated herpesvirus(KSHV), also known as human herpesvirus-8(HHV-8), is etiologically linked to the development of Kaposi’s sarcoma, primary effusion lymphoma, and multicentric Castleman’s disease. These malignancies often occur in immunosuppressed individuals, making KSHV infection-associated diseases an increasing global health concern with persistence of the AIDS epidemic. KSHV exhibits biphasic life cycles between latent and lytic infection and extensive transcriptional and posttranscriptional regulation of gene expression. As a member of the herpesvirus family, KSHV has evolved many strategies to evade the host immune response, which help the virus establish a successful lifelong infection. In this review, we summarize the current research status on the biology of latent and lytic viral infection, the regulation of viral life cycles and the related pathogenesis.展开更多
文摘目的检测EGFR基因在新疆卡波西肉瘤（Kaposi’s Sarcoma，KS）中的表达并分析其与KS发生发展的关系。方法应用SYBR Green I作为荧光染料，以GAPDH作内参照，建立检测EGFR mRNA表达的实时荧光PCR反应体系，对经病理活检证实的7例KS肿瘤组织和瘤旁正常组织中EGFR mRNA的表达水平进行检测。结果KS肿瘤组织中EGFR表达水平低于瘤旁正常组织（t=-3．865，P〈0．05），并且结果有统计学意义。KS肿瘤组织中EGFR的相对表达量为0．34±0．31，瘤旁组织中为1．24±0．78（P=0．008）。结论所建立的SYBR Green Ⅰ实时定量PCR方法可以成功地检测EGFR基因的表达量。KS肿瘤组织中EGFR表达水平低于瘤旁正常组织，差异有统计学意义。
基金Supported by Grant from the National Natural Science Foundation of China(30100160,30271179)
文摘Objective: To study and confirm that recombinant cytokines similar to those produced by HIV-1 infected T cells induced lytic cycle replication of human herpesvirus 8 (HHV-8) in BC-3 cells, another cell line from primary effusion lymphama(PEL). Methods: The persistent stimulation of BC-3 was conducted by several cytokines known to be produced by HIV-1-infected T cells and important in growth and proliferation of Kaposi's sarcoma(KS)cells in vitro, such as the interferon-γ (IFN-γ) , tlie hepatocyte growth factor/scatter factor (HGF / SF) , the Oncostain M(OSM) , and the tumor necrosis factor-α (TNF-α)which is not produced by HIV-1-infected T cells. Treated and untreated BC-3 cells were collected at the 3rd and 7th day of persistent stimulation, respectively. Immuno-histochemical (IHC) staining, Northern blot, quantitative PCR (real- time PCR ) and electron microscopy (EM) were carried out to detect the expression of immunogenic protein ORF59, messenger RNA (mRNA) of minor capsid protein ORF26, and the presence of viral particles of HHV-8 from treated and untreated BC-3 cells. Results: It showed that IFN-γ, HGF/SF, OSM, and TNF-α were found to induce an increase in mRNA expression of ORF26 when added individually to BC-3 cells. Particularly, ORF26 expression stimulated with IFN-γ and TNF-α respectively, increased 6. 1 and 2. 5-fold(from real-time PCR results)at the 7th day when compared with untreated BC-3 cells. Meanwhile, about 20% of IFN-γ stimulated BC-3 cells expressed ORF59 at the 7th day as compared with 1. 5% of untreated BC-3 cells when IHC staining was employed. In addition, viral particles of HHV-8 were readily identified in BC-3 cells stimulated with IFN-γ at the 7th day with EM analysis. Conclusion;TNF-α and recombinant cytokines being similar to those produced by HIV- 1 infected T Cells could really induce HHV- 8 lytic cycle replication in BC-3 cells, another cell line of PEL.
基金supported by the National Key R&D Program of China (2016YFA0502100)the Natural Science Foundation for Distinguished Young Scholars (81425017)+1 种基金the National Institutes of Health awarded (7R01AI116442) to K.Lthe Intramural Research Program of NCI/NIH (1ZIASC010357) to ZMZ
文摘Kaposi’s sarcoma-associated herpesvirus(KSHV), also known as human herpesvirus-8(HHV-8), is etiologically linked to the development of Kaposi’s sarcoma, primary effusion lymphoma, and multicentric Castleman’s disease. These malignancies often occur in immunosuppressed individuals, making KSHV infection-associated diseases an increasing global health concern with persistence of the AIDS epidemic. KSHV exhibits biphasic life cycles between latent and lytic infection and extensive transcriptional and posttranscriptional regulation of gene expression. As a member of the herpesvirus family, KSHV has evolved many strategies to evade the host immune response, which help the virus establish a successful lifelong infection. In this review, we summarize the current research status on the biology of latent and lytic viral infection, the regulation of viral life cycles and the related pathogenesis.
