CARCINOGEN ENHANCEMENT OF EBV INDUCED TRANSFORMATION

我们已经证明，化学和物理致癌物可以增强外周 通过EB病毒（EBV）的血淋巴细胞（PBL）转化。 最近，使用克隆的EB病毒DNA片段，Eco R1 J和D，它们是 在潜伏感染EBV的细胞中转录，我们已经证明， 致癌物诱导的EBV对PBL转化的增强与 EB病毒基因组扩增及其与高分子量 类淋巴母细胞系（LCL）DNA。 这些研究结果一致 结果发现，用病毒产生细胞P3 HR-1处理 N-甲基-N' -亚硝基-亚硝基胍（MNNG）使细胞增殖3倍 在EBV基因组拷贝/细胞和用MNNG预处理Raji细胞中 然后用P3 HR-1病毒重复感染，导致35%的增强 EBV-DNA复制 奇怪的是，每一个观察都强烈地 提示职业环境因素与EBV之间存在协同作用 可能导致EBV相关的恶性肿瘤。 总的来说，这些 观察结果为进一步调查提供了令人信服的论据， 致癌物增强EB病毒诱导淋巴细胞机制 转化，如果这种现象有助于恶性肿瘤， EB病毒感染。 已经从EBV感染的对照中建立了许多LCL， 致癌物处理的PBL，后者具有扩增的EBV序列 并提高生长和克隆效率。 我们建议，通过使用 这些LCL检查EBV基因组的组织及其 通过核酸杂交与宿主细胞基因组的相互作用，我们可以 证明这些参数的某些方面之间的相关性， 加强PBL转化。 具体来说，我们的目标是：1）确定 来自对照的LCL中EBV基因组的甲基化模式 和致癌物处理的PBL; 2）确定EBV序列是否更多 经常发现整合到宿主细胞DNA中的LCL来源于 致癌物处理的PBL;和3）如果是，确定致癌物-EBV是否 淋巴细胞转化过程中协同作用导致癌基因（c-myc或 Blym）激活。 这些实验应该提供信息的核心， 我们对EB病毒如何引发恶性转化的理解。

We have shown that chemical and physical carcinogens can enhance peripheral blood lymphocyte (PBL) transformation by Epstein-Barr virus (EBV). Recently, using cloned EBV DNA fragments, Eco R1 J and D, which are transcribed in cells latently infected with EBV, we have demonstrated that carcinogen-induced enhancement of PBL transformation by EBV correlates with EBV genome amplification and association with high-molecule-weight lymphoblastoid cell line (LCL) DNA. These latter studies are in agreement with the findings that treatment of virus-producing cells P3HR-1 with N-methyl-N' -nitroso-nitrosoguanidine (MNNG) resulted in a 3-fold increase in EBV genome copies per cell and that pretreatment of Raji cells with MNNG followed by superinfection with P3HR-1 virus resulted in a 35% enhancement of EBV-DNA replication. Singularly, each of these observations strongly suggests that synergism between occupational-environmental factors and EBV could contribute to EBV-associated malignancy. Collectively, these observations present a compelling argument for further investigations into the mechanism by which carcinogens enhance EBV-induced lymphocyte transformation and if this phenomenon contributes to malignancy following EBV infection. Numerous LCLs have been established from EBV-infected control and carcinogen-treated PBLs, the latter of which have amplified EBV sequences and enhanced growth and cloning efficiencies. We propose that by using these LCLs to examine the organization of the EBV genome and its interaction with the host cell genome by nucleic acid hybridization, we can demonstrate correlations between certain aspects of these parameters and enhanced PBL transformation. Specifically, our aims are 1) to determine the methylation patterns of the EBV genomes in LCLs derived from control and carcinogen-treated PBLs; 2) to determine whether EBV sequences are more frequently found integrated into host cell DNA in LCLs derived from carcinogen-treated PBLs; and 3) if so, to determine whether carcinogen-EBV synergism during lymphocyte transformation results in oncogene (c-myc or Blym) activation. These experiments should provide information central to our understanding of how the EBV initiates malignant transformation.