IS-RT-PCR ASSAY DETECTION OF MT-MMP IN A HUMAN BREAST-CANCER CELL-LINE

The in situ-reverse transcription-polymerase chain reaction (IS-RT-PCR ) is a method that allows the direct localisation of gene expression. The method utilises the dual buffer mediated activity of the enzyme rT th DNA polymerase enabling both reverse transcription and DNA amplific ation. Labelled nucleoside triphosphates allow the site of expression to be labelled, rather than the PCR primers themselves, giving a more accurate localisation of transcript expression and decreased backgroun d than standard in situ hybridisation (ISH) assays. The MDA-MB-231 hum an breast carcinoma (HBC) cell line was assayed via the IS-RT-PCR tech nique, using primers encoding MT-MMP (membrane-type matrix metalloprot einase) and human beta-actin. Our results clearly indicate baseline ex pression of MT-MMP in the relatively invasive MDA-MB-231 cell line at a signal intensity similar to the housekeeping gene beta-actin, and re sults following induction with Concanavalin A (Con A) are consistent w ith our previous results obtained via Northern blotting.