Raab-Traub N, Hood R, Yang C S, Henry B, Pagano J S
J Virol. 1983 Dec;48(3):580-90. doi: 10.1128/JVI.48.3.580-590.1983.
Sequences which encode Epstein-Barr virus (EBV) RNA in nasopharyngeal carcinoma (NPC) tissue have been identified. We utilized human biopsy material directly as well as NPC grown in nude mice. Total RNA was extracted from the tumor material and separated into polyadenylated and nonpolyadenylated fractions by oligodeoxythymidylate-cellulose chromatography. This material was used as template to construct 32P-labeled cDNA. The labeled cDNAs were hybridized to Southern blots of recombinant EBV DNA fragments. Three of the biopsies, F, 49, and 55, contained polyadenylated RNA homologous to the EBV BamHI fragments V and K, and EcoRI-DIJhet. These same fragments encode the most abundant polyribosomal RNAs in latently infected lymphoblastoid cell lines. The sequences which encoded nonpolyadenylated RNA in NPC tumor 49 were more extensive and included BamHI fragments C, V, B, E, and K, and EcoRI fragments DIJhet, E, F, and G1, a result that indicates selective polyadenylation in EBV RNA processing. A fourth biopsy, NPC tumor 18, contained polyadenylated RNA homologous to the BamHI fragments H, B, K, Y, B1, I1, and A and EcoRI fragments F and G2. A similar pattern of transcription was identified in three tumor specimens from nude mice, 4, 5, and 8. Transformation of lymphocytes did not occur after cocultivation in vitro with explants from these nude mice tumors. This transcriptional pattern may represent an activated state of the EBV genome, formerly not detected in tumor tissue, which is analogous to the state of abortive infection identified in induced in vitro cell systems.
在鼻咽癌(NPC)组织中已鉴定出编码爱泼斯坦-巴尔病毒(EBV)RNA的序列。我们直接利用人体活检材料以及在裸鼠体内生长的NPC。从肿瘤材料中提取总RNA,并通过寡聚脱氧胸苷酸-纤维素色谱法将其分离为聚腺苷酸化和非聚腺苷酸化部分。该材料用作模板构建32P标记的cDNA。将标记的cDNA与重组EBV DNA片段的Southern印迹杂交。活检样本F、49和55中的三个含有与EBV BamHI片段V和K以及EcoRI-DIJhet同源的聚腺苷酸化RNA。这些相同的片段编码潜伏感染的淋巴母细胞系中最丰富的多核糖体RNA。在NPC肿瘤49中编码非聚腺苷酸化RNA的序列更广泛,包括BamHI片段C、V、B、E和K以及EcoRI片段DIJhet、E、F和G1,这一结果表明在EBV RNA加工过程中存在选择性聚腺苷酸化。第四个活检样本NPC肿瘤18含有与BamHI片段H、B、K、Y、B1、I1和A以及EcoRI片段F和G2同源的聚腺苷酸化RNA。在来自裸鼠的三个肿瘤标本4、5和8中鉴定出了类似的转录模式。将淋巴细胞与这些裸鼠肿瘤的外植体在体外共培养后未发生转化。这种转录模式可能代表EBV基因组的一种激活状态,以前在肿瘤组织中未检测到,这类似于在体外诱导细胞系统中鉴定出的流产感染状态。