作者机构：
[甘润良] Pathophysiology Department/Cancer Research Institute,Medical college,Nanhua University,Hengyang
会议名称：
中国病理生理学会第十三届肿瘤、第十四届免疫专业委员会学术会议
会议时间：
2012-06-14
会议地点：
中国河南郑州
摘要：
<正>Background:Epstain-Barr virus（EBV） can transform human B lymphocytes making them immortalized and inducing tumorigenic ability in vitro,but the molecular mechanisms remain unclear.The aim of the present study is to detect and analyze differentially expressed genes in two types of host cells,normal human lymphocytes and coupled EBV-transformed lymphoblasts in vitro using gene chips,and to screen the key regulatory genes of lymphocyte transformation induced by EB virus. Methods:Fresh peripheral blood samples from seven healthy donors were collected. EBV was used to transform lymphocytes in vitro.Total RNA was extracted from 7 cases of the normal lymphocytes and transformed lymphoblasts respectively,marked
作者机构：
[Dai, Yongming] Third People Hosp, Clin Lab, Kunshan City 215300, Jiangsu, Peoples R China.;[He, Fei] Shanghai Ctr Bioinformat Technol SCBIT, Shanghai 200235, Peoples R China.;Univ S China, Canc Res Inst, Sch Med, Hengyang City 421001, Hunan, Peoples R China.;[Wu, Yimou] Univ S China, Pathogen Biol Inst, Hengyang 421001, Peoples R China.
通讯机构：
[Runliang Gan] C;[Yimou Wu] P;Cancer Research Institute, University of South China, People’s Republic of China<&wdkj&>Cancer Research Institute, University of South China, School of Medicine, Hengyang City, People’s Republic of China<&wdkj&>Pathogenic Biology Institute, University of South China, People’s Republic of China<&wdkj&>Pathogenic Biology Institute, University of South China, Hengyang, China
关键词：
Epstein-Barr virus (EBV);Lymphocyte transformation;Lymphoblastoid cell line (LCL);Gene expression;Gene chip
摘要：
Epstain-Barr virus (EBV) can transform human B lymphocytes making them immortalized and inducing tumorigenic ability in vitro, but the molecular mechanisms remain unclear. The aim of the present study is to detect and analyze differentially expressed genes in two types of host cells, normal human lymphocytes and coupled EBV-transformed lymphoblasts in vitro using gene chips, and to screen the key regulatory genes of lymphocyte transformation induced by EB virus. Fresh peripheral blood samples from seven healthy donors were collected. EBV was used to transform lymphocytes in vitro. Total RNA was extracted from 7 cases of the normal lymphocytes and transformed lymphoblasts respectively, marked with dihydroxyfluorane after reverse transcription, then hybridized with 4 × 44 K Agilent human whole genome microarray. LIMMA, String, Cytoscape and other softwares were used to screen and analyze differentially expressed genes. Real-time PCR was applied to verify the result of gene expression microarrays. There were 1745 differentially expressed genes that had been screened, including 917 up-regulated genes and 828 down-regulated genes. According to the results of Generank, String and Cytoscape analyses, 38 genes may be key controlled genes related to EBV-transformed lymphocytes, including 22 up-regulated genes(PLK1, E2F1, AURKB, CDK2, PLCG2, CD80, PIK3R3, CDC20, CDC6, AURKA, CENPA, BUB1B, NUP37, MAD2L1, BIRC5, CDC25A, CCNB1, RPA3, HJURP, KIF2C, CDK1, CDCA8) and 16 down-regulated genes(FYN, CD3D, CD4, CD3G, ZAP70, FOS, HCK, CD247, PRKCQ, ITK, LCP2, CXCL1, CD8A, ITGB5, VAV3, CXCR4), which primarily control biological processes such as cell cycle, mitosis, cytokine-cytokine pathway, immunity response and so on. Human lymphocyte transformation induced by EB virus is a complicated process, involving multiple-genes and –pathways in virus-host interactions. Global gene expression profile analysis showed that EBV may transform human B lymphocytes by promoting cell cycle and mitosis, inhibiting cell apoptosis, hindering host immune function and secretion of cytokines.
作者机构：
[Gan, Runliang; Zhang, Yang; Wu, Yimou; Cheng, Ailan; Tang, Yunlian; Liu, Fang] Univ S China, Canc Res Inst, Hengyang City 421001, Hunan, Peoples R China.;[He, Rongfang] Univ S China, Dept Pathol, Affiliated Hosp 1, Hengyang 421001, Hunan, Peoples R China.
通讯机构：
[Gan, Runliang] U;Univ S China, Canc Res Inst, Hengyang City 421001, Hunan, Peoples R China.
关键词：
SCID Mouse;Lymphoproliferative Disease;Human Peripheral Blood Lymphocyte;Leukocyte Common Antigen;EBER1 Probe
摘要：
Epstein-Barr virus (EBV) has a close association with various types of human lymphomas. Animal models are essential to elucidate the pathogenesis of human EBV-associated lymphomas. The aim of the present study is to evaluate the association between human IgG concentration and EBV-associated lymphoma development in huPBL/SCID mice. Human peripheral blood lymphocytes (hu-PBL) from EBV-seropositive donors were inoculated intraperitoneally into SCID mouse. Immunohistochemical staining was used to examine differentiated antigens of tumor cells. EBV infection of the induced tumors was detected by in situ hybridization. IgG concentrations in the serums of 12 SCID mice were measured by unidirectional immunodiffusion assay. 21 out of 29 mice developed tumors in their body. Immunohistochemical staining showed that all induced tumors were LCA (leukocyte common antigen) positive, B-cell markers (CD20, CD79a) positive, and T-cell markers (both CD3 and CD45RO) negative. The tumors can be diagnosed as human B-cell lymphomas by these morphological and immunohistochemical features. In situ hybridization exhibited resultant tumor cells had EBV encoded small RNA-1 (EBER-1). Human-derived IgG could be found in the serum from SCID mice on the 15th day following hu-PBL transplantation, and IgG levels increased with the tumor development in 6 hu-PBL/SCID chimeras. Intraperitoneal transfer of hu-PBLs from EBV+ donors to SCID mice leads to high human IgG levels in mouse serum and B cell lymphomas. Our findings suggest that increasing levels of human-derived IgG in peripheral blood from hu-PBL/SCID mice could be used to monitor EBV-related human B-cell lymphoma development in experimental animals.
摘要：
EB病毒（EBV）是淋巴瘤发生的一个重要因素,EB病毒基因表达产物潜伏膜蛋白（1atentmembrane protein LMP）包括LMP-1、LMP-2A和LMP-2B。LMP-1通常被认为是病毒致瘤基因,是肿瘤形成过程中的主要促进因子。近年来研究表明LMP-1具有介导细胞增殖、抑制细胞凋亡的功能。采集健康献血员的外周静脉血,分离正常人淋巴细胞,移植到严重联合免疫缺陷动物SCID小鼠体内,建立SCID小鼠体内EBV感染诱发淋巴瘤的动物模型。病理学检查确定诱发瘤是人缘性B细胞淋巴瘤。应用实时定量PCR方法检测正常人淋巴细胞、诱发淋巴瘤细胞中Lmp（LMP-1、LMP-2A、LMP-2B）的差异表达情况。实时定量PCR结果表明:LMP-1、LMP-2A在诱发淋巴瘤细胞中均表达上调,LMP-2B在"正常人"淋巴细胞、诱发淋巴瘤细胞中均表达。Western-blot检测LMP-1在诱发淋巴瘤细胞中表达比在正常人淋巴细胞中增强,这些结果提示在EBV诱导淋巴瘤发生过程中LMP可能起重要作用。
