Kassem Ahmad, Schöpflin Anja, Diaz Carlos, Weyers Wolfgang, Stickeler Elmar, Werner Martin, Zur Hausen Axel
Institute of Pathology, University Hospital Freiburg, and Center for Dermatopathology, Freiburg, Germany.
Cancer Res. 2008 Jul 1;68(13):5009-13. doi: 10.1158/0008-5472.CAN-08-0949.
Merkel cell carcinoma (MCC) is a rare but very aggressive human malignancy of the elderly or immunosuppressed patients. Recently, the clonal integration of a new human polyoma virus, which was termed Merkel cell polyomavirus (MCPyV), has been reported in 8 of 10 MCC patients. In the present study, we studied the formalin-fixed and paraffin-embedded tissue specimens of 39 MCC for the presence of MCPyV by PCR. We applied four different primer sets directed against the large T antigen and the VP1 gene of MCPyV. We were able to detect MCPyV in 77% (n = 30) of MCC as confirmed by sequence analyses of the PCR products. Sequence analyses showed only minor nucleotide changes compared with the previously published MCC sequences. In addition, one patient revealed the amplification of two PCR products using PCR primers directed against the VP1 gene. Sequence analyses confirmed the presence of the expected 351-bp PCR product and in addition a second PCR product of 261 bp containing a unique 90-bp deletion in the VP1 gene, which will lead to a predicted loss of 28 amino acids. The unique 90-bp deletion within the VP1 gene possibly is a result of incomplete viral integration of MCPyV in the MCC. The presence of MCPyV in the majority of MCC tissue specimens in our study strongly underlines a possible role for MCPyV as an etiologic agent in the carcinogenesis of MCC.
默克尔细胞癌(MCC)是一种罕见但侵袭性很强的人类恶性肿瘤,多见于老年人或免疫抑制患者。最近,在10例MCC患者中有8例报告了一种新的人类多瘤病毒——默克尔细胞多瘤病毒(MCPyV)的克隆整合。在本研究中,我们通过聚合酶链反应(PCR)检测了39例MCC福尔马林固定石蜡包埋组织标本中MCPyV的存在情况。我们应用了四种不同的引物组,分别针对MCPyV的大T抗原和VP1基因。通过对PCR产物的序列分析证实,我们在77%(n = 30)的MCC中检测到了MCPyV。序列分析显示,与先前发表的MCC序列相比,只有微小的核苷酸变化。此外,一名患者使用针对VP1基因的PCR引物扩增出了两种PCR产物。序列分析证实了预期的351 bp PCR产物的存在,此外还证实了一个261 bp的第二种PCR产物的存在,该产物在VP1基因中含有一个独特的90 bp缺失,这将导致预测的28个氨基酸的缺失。VP1基因内独特的90 bp缺失可能是MCPyV在MCC中病毒整合不完全的结果。我们研究中大多数MCC组织标本中存在MCPyV,这有力地强调了MCPyV作为MCC致癌病因的可能作用。