Development and primary application of a real-time quantitative polymerase chain reaction for the detection of human breast cancer related novel gene-kallikrein gene 6 expression / 中华检验医学杂志

ABSTRACT

Objective To develop a real-time quantitative PCR method for detection of human breast cancer related novel gene-kallikrein gene 6 (klk6) expression and investigate klk6 expression levels in breast cancer tissue.Methods Using Sybr Green I, with GAPDH as reference, a real-time quantitative PCR method was established. klk6 expression levels of 32 normal and breast cancer tissues were detected and analyzed by the method and REST software.Results The amplification efficiencies for GAPDH and klk6 of the real-time quantitative PCR method were 0.90 And 0.95, respectively; inter-coefficient of variation were 1.0%~2.1% and 0.8%~1.2%,respectively; intra-coefficient of variation were 3.2% and 3.9%, respectively. The relative expression levels of klk6 in normal breast and breast cancer tissues were 0.017?0.009 and 0.040?0.017 with GAPDH as reference. Analysis results with REST indicated klk6 expression was up- regulated in breast cancer.Conclusion The real-time quantitative PCR method with Sybr Green I for klk6 mRNA quantification was simple, specific, reproductive and reliable, and could be used to study relationship betweeen klk6 expression and tumor.