Prevalence of Merkel cell polyomavirus in Merkel cell carcinoma
Prevalence of Merkel cell polyomavirus in Merkel cell carcinoma
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DOI：
10.1038/modpathol.2009.3
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发表时间：
2009-04-01
期刊：
影响因子：
7.5
通讯作者：
Pfeifer, John D.
中科院分区：
文献类型：
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作者：
Duncavage, Eric J.;Zehnbauer, Barbara A.;Pfeifer, John D.
最近有研究表明，默克尔细胞癌是一种罕见的、往往是致命的皮肤恶性肿瘤，它经常携带一种新的克隆性整合多瘤病毒，恰如其分地命名为默克尔细胞多瘤病毒。我们的目的是研究默克尔细胞多瘤病毒在默克尔细胞癌病例中的流行情况，使用福尔马林固定的石蜡包埋组织块的标本。在我们的档案中，我们确定了41例默克尔细胞癌(来自29个不同的患者)。其中原发皮肤肿瘤20例，局部复发4例，转移17例。用先前发表的两组引物LT1(440bp扩增子)和LT3(308bp扩增子)以及新的引物组MCVPS1(109bp扩增子)对所有病例进行了聚合酶链式反应。对选定的聚合酶链式反应产物进行测序，以确认扩增子的特性。此外，用BamH1对MCVPS1产物进行酶切，得到83个碱基的产物。所有41例研究病例均可扩增出DNA。三对引物的MERKEL细胞多瘤病毒的检出率分别为：MCVPS1患者22例(76%)，LT3患者12例(41%)，LT1患者8例(28%)。引物组检测率的差异在很大程度上是由于DNA质量较差，支持这一点的是大小控制梯形产物中较高分子量标记的扩增不佳，以及MCVPS1对LT1和LT3阳性的所有病例都是阳性的。我们的发现提供了进一步的证据，证明默克尔细胞多瘤病毒与默克尔细胞癌的发生有关。在更实际的层面上，我们的石蜡优化的引物集可以用作辅助测试，以在临床环境中确认默克尔细胞癌的诊断，或用于筛查其他罕见的肿瘤类型的致病病毒，特别是那些在冷冻组织库中代表性不足的肿瘤类型。
It has recently been shown that Merkel cell carcinoma, a rare and often lethal cutaneous malignancy, frequently harbors a novel clonally integrated polyomavirus aptly named Merkel cell polyomavirus. We aimed to study the prevalence of Merkel cell polyomavirus in cases of Merkel cell carcinoma, using specimens from formalin-fixed, paraffin- embedded tissue blocks. In our archives we identified 41 cases of Merkel cell carcinoma (from 29 different patients). Of these, 20 cases were primary cutaneous tumors, 4 were local recurrences, and 17 were metastases. PCR using two previously published primer sets, LT1 (440 bp amplicon) and LT3 (308 bp amplicon), as well as a novel primer set MCVPS1 (109 bp amplicon), was performed on all cases. Selected PCR products were sequenced to confirm amplicon identity. In addition, the MCVPS1 products were digested with BamH1, yielding an 83 bp product. Amplifiable DNA was recovered in all 41 study cases. The detection rate of Merkel cell polyomavirus for each of the three primer sets was 22 of 29 patients (76%) for MCVPS1, 12 of 29 (41%) for LT3, and 8 of 29 (28%) for LT1. The variation between primer set detection rates was largely due to poor DNA quality, as supported by poor amplification of the higher molecular weight markers in size control ladder products and the fact that all cases that were positive by LT1 and LT3 were positive by MCVPS1. Our findings provide further evidence to link Merkel cell polyomavirus with a possible role in the oncogenesis of Merkel cell carcinoma. On a more practical level, our paraffin-optimized primer set may be used as an ancillary test to confirm the diagnosis of Merkel cell carcinoma in the clinical setting or for screening other rare tumor types for the causative virus, especially those tumor types that are underrepresented in frozen tissue repositories.