Mutiu A, Alexiu I, Chivu M, Petica M, Anton G, Bleotu C, Diaconu C, Popescu C, Jucu V, Cernescu C
St. S. Nicolau Institute of Virology, 285 sos Mihai Bravu, Bucharest 79650, Romania.
J Cell Mol Med. 2001 Jan-Mar;5(1):49-59. doi: 10.1111/j.1582-4934.2001.tb00137.x.
The role of Human Papillomaviruses (HPV) in laryngeal carcinomas has been studied with conflicting results. To evaluate the etiologic relationship between HPV infection and epithelial malignancy of the larynx we studied five laryngeal carcinoma cell lines obtained from patients undergoing surgery for laryngeal tumors. The paraffin embedded biopsy samples of the original tumor and different passages of the new established cell lines were investigated by PCR with consensus primers specific for HPV DNA. The findings indicate that HPV infection is associated with some larynx carcinomas. The positive association has been enhanced when a method of enrichment of epithelial cells from fresh tumor samples was used. All tumor cells enriched smears were positive for HPV DNA not only by PCR but also by in situ hybridization (ISH). Investigated by PCR, different passages of larynx tumor cell lines maintained expression of HPV DNA. At subsequent passages ISH gives constantly no signals suggesting a minimal amount of viral harbored sequences. In one cell line propagated more than 60 population doublings, the chromosomal frequency distribution shifted from modal number 46 at the 5(th) passage to 63 at the 60(th) passage. The mechanisms by which persistent HPV infection maintains continuous cell proliferation were discussed.
人乳头瘤病毒(HPV)在喉癌中的作用已得到研究,但结果相互矛盾。为了评估HPV感染与喉上皮恶性肿瘤之间的病因关系,我们研究了从接受喉肿瘤手术的患者中获取的5种喉癌细胞系。用针对HPV DNA的共有引物通过聚合酶链反应(PCR)对原肿瘤的石蜡包埋活检样本以及新建立的细胞系的不同传代样本进行了研究。研究结果表明,HPV感染与某些喉癌有关。当使用从新鲜肿瘤样本中富集上皮细胞的方法时,这种正相关关系得到了加强。所有富集肿瘤细胞的涂片不仅通过PCR检测HPV DNA呈阳性,而且通过原位杂交(ISH)检测也呈阳性。通过PCR检测发现,喉肿瘤细胞系的不同传代均保持HPV DNA的表达。在随后的传代中,ISH持续未给出信号,提示病毒携带序列的量极少。在一个传代超过60次群体倍增的细胞系中,染色体频率分布从第5代时的众数46条转变为第60代时的63条。文中讨论了持续性HPV感染维持细胞持续增殖的机制。