Lymphangiogenesis quantification using quantitative PCR and breast cancer as a model

The detection of lymphangiogenesis (formation of new lymphatics) has previo usly been difficult to measure, primarily due to the lack of specific marke rs for lymphatic endothelium. Using conventional PCR (polymerase chain reac tion), DNA sequencing, plasmid synthesis, and real-time quantitative PCR (R TQPCR), we report a new approach to enable the measurement of lymphangiogen esis using LYVE-1, a novel, specific lymphatic marker in breast cancer tiss ue. By using a Scorpion-based probe system with the RTQPCR analyser, a high ly sensitive and specific detection and quantitation of LYVE-1 was possible . It was found that lymphangiogenesis occurred in all breast specimens and that higher levels were found in tumours which had spread to the lymph node s. (C) 2001 Academic Press.