Detection of soluble Fas mRNA using in situ reverse transcription- polymerase chain reaction

Fas protein (Fas) is known to induce cell death by apoptosis in susceptible cells. Alternative splicing of the Fas gene produces soluble Fas protein (sFas), which is considered to block the function of Fas. The serum level of sfas is elevated in patients with various malignancies in a manner reflective of disease stage and tumor burden, but the precise cellular origin of sfas in vivo has not yet been clarified. To identify the cells that synthesize sfas mRNA on histologic specimens, we applied in situ reverse transcription-polymerase chain reaction (in situ RT-PCR) in 11 cases of gastric adenocarcinoma/metastatic lymph node. Furthermore, we studied the distribution of Fas using immunohistochemistry and Fas mRNA using in situ RT- PCR. In all primary tumors and 10 of 11 metastatic tumors, tumor cells expressed both Fas- and sfas mRNA. Lymphocytes infiltrated in the tumor tissues and the lymph nodes also revealed both mRNA signals. A clear correlation between the tissue distribution for Fas and its mRNA was also observed. These observations demonstrated that solid tumors in vivo can synthesize sfas mRNA and suggest that tumor cells are responsible in part for elevated sfas in human malignancies. However, the additional expression of sfas mRNA in tissue lymphocytes indicates the complex regulatory mechanisms of Fas-mediated apoptosis pathway in tumor pathogenesis and host defense. We also demonstrated that in situ RT-PCR can be a suitable method for in situ detection of alternatively spliced mRNA.