Role of the Novel Nuclear Tyrosine Kinase, RAK, in Breast Cancer Biology

摘要

The research supported by this grant is intended to investigate the feasibility of an electrocatalytic guanine oxidation reaction as an mRNA quantification method. This system is to be tested first by quantifying the mRNA of Rak nuclear tyrosine kinase in both cell culture and breast tissue samples. To accomplish this goal a competitive RT-PCR assay was designed to measure the absolute quantity of Rak mRNA in each sample. This absolute quantity was compared to the electrochemical signal generated by oxidation of the Rak RT-PCR products immobilized to the indium tin oxide (ITO) electrodes used in this study. This final report presents data that show Rak to be overexpressed in 33% of the breast cancers analyzed. Rak mRNA in normal breast tissue is expressed at the level of roughly 400 zmol/ug total RNA; however, Rak was overexpressed to the level of roughly 2 amol/ug total RNA in some breast tumors analyzed. The breast tumors that exhibited this high level of overexpression by competitive RT-PCR also exhibited the largest electrochemical signal (current) when catalytic guanine oxidation was used as the detection and quantification method.