Quantitative real-time reverse transcription-PCR assay for urokinase plasminogen activator, @entity2289 , and XXXX gene expressions in primary @entity0 .

@entity1 @entity217 @entity2289 @entity0 @entity5 @entity2843 @entity3 @entity2290
BACKGROUND: The plasminogen activation system and matrix metalloproteinases (MMPs) play a key role in the degradation of basement membrane and extracellular matrix in tissue remodeling, @entity5 cell invasion, and @entity3 . METHODS: Quantitative real-time reverse-transcription-PCR (RT-PCR) assays were developed to quantify @entity2290 ( @entity2290 ), @entity2289 ( @entity2289 ), and @entity2843 ( @entity2843 ) mRNA in 54 @entity0 tissues. Gene fragments were amplified in a LightCycler real-time PCR system using gene-specific primers and SYBR Green I. The results were normalized to @entity217 mRNA. We also quantified antigen and functional concentrations of these components. RESULTS: The intra- and interassay variabilities for mRNA quantification showed mean SDs for the crossing point of 0.12 and 0.15 cycles, respectively. @entity2289 , @entity2290 , and @entity2843 mRNA and antigen concentrations and @entity2289 and @entity2290 functional concentrations increased with @entity5 severity; the increase was statistically significant for @entity2289 , @entity2290 , and @entity2843 mRNA and antigen concentrations and for @entity2290 functional concentrations. Node-positive @entity1 showed significantly higher @entity2289 , @entity2290 , and @entity2843 mRNA and antigen concentrations than those who were node negative. CONCLUSIONS: Quantitative real-time RT-PCR is a highly sensitive, reproducible, and fast method for measuring gene expression of @entity2289 , @entity2290 , and @entity2843 in @entity0 . These components may be involved in @entity0 development, and increased mRNA expression may be associated with a worse prognosis.
