ABSTRACT
The RON gene, encoding the tyrosine-kinase receptor for macrophage-stimulating protein (MSP), is involved in a range of neoplastic processes. However, no aberration in RON or MSP has been identified in Merkel cell carcinoma (MCC). We investigated the RON signaling pathway in MCC. Fourteen cases of MCC were tested for the expression of RON and its ligand, MSP, using reverse transcription PCR (RT-PCR) and immunohistochemistry. The mutation of RON was also examined. RT-PCR identified transcription of both RON and MSP in all nine cases that were available for the examination. Immunohistochemistry showed the expression of RON (9/14 cases) and MSP (9/14 cases). Six cases out of 14 were positive for both RON and MSP. Normal Merkel cells were negative for RON and MSP expression. Missense mutation in the tyrosine kinase region of RON was detected in one of the 14 cases, and the case showed expression of RON. Transcription of RON and MSP was observed in MCC, and a considerable number of MCC cases expressed both RON and MSP, while Merkel cells do not express these molecules. The results suggest that RON signaling seems to play at least some role in the pathogenesis of MCC.
Subject(s)
Carcinoma, Merkel Cell/metabolism , Hepatocyte Growth Factor/metabolism , Proto-Oncogene Proteins/metabolism , Receptor Protein-Tyrosine Kinases/metabolism , Skin Neoplasms/metabolism , Aged , Aged, 80 and over , Base Sequence , Carcinoma, Merkel Cell/genetics , Carcinoma, Merkel Cell/pathology , Female , Humans , Immunohistochemistry , Laser Capture Microdissection , Male , Middle Aged , Mutation, Missense , Polymorphism, Single-Stranded Conformational , Receptor Protein-Tyrosine Kinases/genetics , Signal Transduction/physiology , Skin Neoplasms/genetics , Skin Neoplasms/pathologyABSTRACT
OBJECTIVE: To apply the polymerase chain reaction (PCR) for detection of the HTLV-I gene from cytologic smear slides. STUDY DESIGN: Samples were from seven cases of serum anti-ATL antibody (ATLA)-positive T-cell lymphoma and three from ATLA-negative T-cell lymphoma. Six of the seven ATLA-positive cases were confirmed to be ATLL by Southern blotting. From the seventh case a fresh sample for blotting could not obtained. DNA was extracted from the cytologic smear slides of all 10 cases; they had been stained with Papanicolaou or May-Giemsa stain, digested with proteinase K and precipitated with phenol and ethanol. The target sequence in the pX region of the HTLV-I gene was amplified by PCR. RESULTS: All seven ATLA-positive cases, including one that had not yet been confirmed by Southern blotting, showed a single band, as predicted, while the three ATLA-negative cases showed no band. CONCLUSION: If cytologic smear slides are available but a fresh sample is not, the PCR method should provide evidence that the virus is present since in our study sufficient DNA templates were successfully extracted from the stained cytologic smear slides for detection of the virus.
