Lymphangiogenesis quantification using quantitative PCR and breast cancer as a model.

The detection of lymphangiogenesis (formation of new lymphatics) has previously been difficult to measure, primarily due to the lack of specific markers for lymphatic endothelium. Using conventional PCR (polymerase chain reaction), DNA sequencing, plasmid synthesis, and real-time quantitative PCR (RTQPCR), we report a new approach to enable the measurement of lymphangiogenesis using LYVE-1, a novel, specific lymphatic marker in breast cancer tissue. By using a Scorpion-based probe system with the RTQPCR analyser, a highly 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 nodes. Copyright 2001 Academic Press.