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Fluorescence in situ hybridization and qPCR to detect Merkel cell polyomavirus physical status and load in Merkel cell carcinomas
Author(s) -
Haugg Anke M.,
Rennspiess Dorit,
Hausen Axel zur,
Speel ErnstJan M.,
Cathomas Gieri,
Becker Jürgen C.,
Schrama David
Publication year - 2014
Publication title -
international journal of cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.475
H-Index - 234
eISSN - 1097-0215
pISSN - 0020-7136
DOI - 10.1002/ijc.28931
Subject(s) - merkel cell polyomavirus , fluorescence in situ hybridization , merkel cell carcinoma , biology , in situ hybridization , merkel cell , carcinogenesis , tissue microarray , virus , viral load , context (archaeology) , virology , pathology , cancer , carcinoma , gene expression , genetics , medicine , gene , chromosome , paleontology
The Merkel cell polyomavirus (MCPyV) is detected in 80% of Merkel cell carcinomas (MCC). Clonal integration and tumor‐specific mutations in the large T antigen are strong arguments that MCPyV is a human tumor virus. However, the relationship between viral presence and cancer induction remains discussed controversially. Since almost all studies on virus prevalence are based on PCR techniques, we performed MCPyV fluorescence in situ hybridization (FISH) on MCC to gain information about the quality of the viral presence on the single cell level. MCPyV‐FISH was performed on tissue microarrays containing 62 formalin‐fixed and paraffin‐embedded tissue samples including all tumor grades of 42 patients. The hybridization patterns were correlated to the qPCR data determined on corresponding whole tissue sections. Indeed, MCPyV‐FISH and qPCR data were highly correlated, i.e . 83% for FISH‐positive and 93% for FISH‐negative cores. Accordingly, the mean of the qPCR values of all MCPyV‐positive cores differed significantly from the mean of the negative cores ( p = 0.0076). Importantly, two hybridization patterns were definable in the MCPyV‐FISH: a punctate pattern (85%) indicating viral integration, which correlated with a moderate viral abundance and a combination of the punctate with a diffuse pattern (15%), suggesting a possible coexistence of integrated and episomal virus which was associated with very high viral load and VP1 expression. Thus, MCPyV‐FISH adds important information on the single cell level within the histomorphological context and could therefore be an important tool to further elucidate MCPyV related carcinogenesis.