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Protoporphyrin IX Occurs Naturally in Colorectal Cancers and Their Metastases¹

Robert-Roessle-Hospital at the Max-Delbrueck-Center for Molecular Medicine, Charité, Humboldt-University at Berlin, Berlin, Germany [K. T. M., T. H., C. N., W. E. H., P. M. S.]; Division of Medical Physics and Metrological Information Technology, Section of Biomedical Optics and NMR-Measuring Techniques, Physikalisch-Technische Bundesanstalt, Berlin, Germany [B. E., D. N., H. R.]; and Division of Radiation Biology, Roswell Park Cancer Institute, Buffalo, New York [R. K. P., T. J. D.]

Colorectal cancers exhibit a red fluorescence. The nature of the responsible fluorophore and its eventual diagnostic potential were investigated. Thirty-three consecutive colorectal resection specimen, 32 of which with histologically confirmed cancer, and a total of 1053 palpable mesenteric nodes were fluorimetrically characterized ex vivo. Furthermore, frozen material from 28 patients was analyzed, selected for the availability of primary tumor material and metastatic tissue, e.g., lymphatic and liver metastases from the same patient. Biochemical characterization was carried out through chemical extraction and reversed phase high-performance liquid chromatography. The fluorescence spectra of tissues, tissue extracts, and standard solutions of porphyrins were determined using a pulsed solid-state laser system for excitation and an imaging polychromator, together with an intensified CCD camera for time-delayed observation. Protoporphyrin IX (PpIX) was identified as the predominant fluorophore in primary tumors and their metastases. The fluorophore occurred in the absence of necrosis and in sterile locations. In untreated cases (n = 24), PpIX fluorescence discriminates metastatically involved lymph nodes from all other palpable nodes with a sensitivity of 62% at a specificity of 78% (P < 0.0001). After neoadjuvant treatment of rectal cancer, the PpIX fluorescence level of the primary tumors was reduced and a discrimination of lymph nodes based on PpIX-fluorescence was impossible. We conclude that colorectal cancer metastases accumulate diagnostic levels of endogenous PpIX as a result of a tumor-specific metabolic alteration.

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