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
- VernacularTitle:乳腺癌klk6基因表达的实时荧光定量法的建立及初步应用
- Author:
Chengjin HU
;
Qian SHEN
;
Yingjian CHEN
;
Xinmian WEN
- Publication Type:Journal Article
- Keywords:
Breast neoplasms;
Reverse transcriptase polyerase chain reaction;
Kallikrein gene
- From:
Chinese Journal of Laboratory Medicine
2000;0(06):-
- CountryChina
- Language:Chinese
-
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.
