Gan Run-Liang, Yin Zhi-Hua, Liu Teng-Fei, Dong Bi-Hua, Zhou Jian-Guo, Yao Kai-Tai
Cancer Research Institute, Nanhua University, Hengyang 421001, China.
Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai). 2003 Oct;35(10):925-9.
To identify molecular features of neoplasms associated with EB virus, human peripheral blood lymphocytes (huPBL) were isolated from healthy volunteer donors and were transplanted intraperitoneally into SCID mice, and then huPBL/SCID mice were infected with EB virus. Serum levels of human IgG were measured by unidirectional immunodiffusion assay. Human Alu sequence and EBER-1 in tumor tissues were detected with PCR and in situ hybridization. Immunohistochemical staining was used to examine leukocyte differentiation antigens (LCA, L26, UCHL1, PS1), viral gene products (LMP1, EBNA2, BZLF1) and cellular oncoproteins (p53, C-myc, Bcl-2 and Bax). The experiments showed that tumors developed in 24 of 34 surviving huPBL/SCID mice by EBV infection. Histopathological and immunohistochemical observations demonstrated that all of the induced tumors in SCID mice were malignant lymphomas derived from human B-lymphocytes. In situ hybridization showed that tumor cells had EBV-encoded small RNA-1 (i.e. EBER-1). Alu sequence could be amplified by PCR from human genome of tumor tissues. Immunohistochemistry detected positive staining of BZLF1-encoded protein in a small population of tumor cells of almost all cases, and positive staining of LMP1 and EBNA2 only in small number of tumor cells. Human IgG could be found in the serum of 12 SCID mice on the 15th day after huPBL engraftment, and then increased with time and with the development of induced tumors in 6 mice. Positive rates of p53, C-myc, Bcl-2 and Bax expression were 83.33%, 100%, 95.83%, 91.67%, respectively, in 24 cases of the EBV-induced lymphomas. The results indicate that molecular lesions associated with the induced B-cell lymphoma involved EBV infection, expression of oncogenic viral genes, and abnormal expression of cellular oncogenes in human xenografts. Human IgG level in the serum of huPBL/SCID mice can be considered as a useful index for tumor development.
为了鉴定与EB病毒相关的肿瘤的分子特征，从健康志愿者供体中分离出人外周血淋巴细胞（huPBL），并将其腹腔内移植到SCID小鼠体内，然后用EB病毒感染huPBL/SCID小鼠。通过单向免疫扩散试验测量人IgG的血清水平。用PCR和原位杂交检测肿瘤组织中的人Alu序列和EBER-1。免疫组织化学染色用于检测白细胞分化抗原（LCA、L26、UCHL1、PS1）、病毒基因产物（LMP1、EBNA2、BZLF1）和细胞癌蛋白（p53、C-myc、Bcl-2和Bax）。实验表明，34只存活的huPBL/SCID小鼠中有24只通过EB病毒感染发生了肿瘤。组织病理学和免疫组织化学观察表明，SCID小鼠中所有诱导的肿瘤均为源自人B淋巴细胞的恶性淋巴瘤。原位杂交显示肿瘤细胞具有EB病毒编码的小RNA-1（即EBER-1）。Alu序列可通过PCR从肿瘤组织的人类基因组中扩增出来。免疫组织化学在几乎所有病例的少数肿瘤细胞中检测到BZLF1编码蛋白的阳性染色，仅在少数肿瘤细胞中检测到LMP1和EBNA2的阳性染色。在huPBL植入后第15天，12只SCID小鼠的血清中可检测到人IgG，然后随着时间的推移以及6只小鼠中诱导肿瘤的发展而升高。在24例EB病毒诱导的淋巴瘤中，p53、C-myc、Bcl-2和Bax表达的阳性率分别为83.33%、100%、95.83%、91.67%。结果表明，与诱导的B细胞淋巴瘤相关的分子损伤涉及EB病毒感染、致癌病毒基因的表达以及人异种移植中细胞癌基因的异常表达。huPBL/SCID小鼠血清中的人IgG水平可被视为肿瘤发展的有用指标。
