Combined application of RT-PCR and immunohistochemistry on paraffin embedded sentinel lymph nodes of prostate cancer patients

• Published in: Pathology, research and practice research and practice, 2005-01, Vol.200 (11), p.763-770
• Main Authors: Haas, Christian J., Wagner, Theodor, Wawroschek, Friedhelm, Arnholdt, Hans
• Format: Article
• Language: English
• Subjects: Adenocarcinoma - genetics; Adenocarcinoma - metabolism; Adenocarcinoma - secondary; Antigens, Surface - genetics; Antigens, Surface - metabolism; Biomarkers, Tumor - genetics; Biomarkers, Tumor - metabolism; Glutamate Carboxypeptidase II - genetics; Glutamate Carboxypeptidase II - metabolism; Humans; Immunoenzyme Techniques - methods; Lymph Nodes - metabolism; Lymph Nodes - pathology; Lymphatic Metastasis; Male; Paraffin embedded tissue; Paraffin Embedding; Predictive Value of Tests; Prostate cancer; Prostate-Specific Antigen - genetics; Prostate-Specific Antigen - metabolism; Prostatic Neoplasms - genetics; Prostatic Neoplasms - metabolism; Prostatic Neoplasms - pathology; PSA; Reverse Transcriptase Polymerase Chain Reaction; RNA, Neoplasm - analysis; RT-PCR; Sentinel lymph node
• ISSN: 0344-0338; 1618-0631
• DOI: 10.1016/j.prp.2004.09.008

The detection of tumor cells in the sentinel lymph node (SLN) is of great importance for the prognosis of cancer patients. At present, immunohistochemistry and RT-PCR for tumor marker expression are the most sensitive techniques available for this analysis. However, so far, most RT-PCR-based analyses of SLNs have been performed on fresh material, excluding a direct comparison with the (immuno)histologic results. In our view, this does not entirely aid routine diagnosis. We established an efficient method for RNA extraction and RT-PCR from paraffin sections of SLNs from prostate cancer patients and compared the results with the (immuno)histologic data of adjacent sections. Amplifiable RNA was obtained from 133 SLNs of 68 prostate cancer patients. Correlation of PSA-specific RT-PCR with (immuno)histologic findings showed a positive and negative predictive value of 83% and 100%, respectively, for the prostate cancer patients investigated. Four of 12 patients with biochemical relapse, but without (immuno)histologically detectable tumor cells were RT-PCR-positive for PSA. We found that single sections of paraffin-embedded SLNs are suitable for routinely performed RT-PCR. Combined with (immuno)histology, PSA-specific RT-PCR is a revealing supplementary technique for the detection of tumor cells in SLNs of prostate cancer patients.