A rapid colorimetric in situ messenger RNA hybridization technique for analysis of epidermal growth factor receptor in paraffin-embedded surgical specimens of human colon carcinomas.

摘要: Abstract We have developed a rapid colorimetric in situ mRNA hybridization procedure to analyze epidermal growth factor receptor (EGF-R) transcripts paraffin-embedded surgical specimens of human colon carcinomas. This technique is based on the use 24-base oligonucleotide probes labeled with 6 biotin molecules at 3′ end. integrity was verified using hyperbiotinylated 30-residue-long deoxythymidylate probe, and specificity reaction confirmed by EGF-R-specific sense antisense probes. Avidin alkaline phosphatase detection capillary technology used Microprobe System allowed for completion under 5 h. The A431 epidermoid carcinoma cells growing culture fixed formalin as well sections this tumor s.c. nude mice served positive controls. In EGF-R directly correlated protein levels observed Northern blot immunohistochemistry, respectively. primary metastases from liver lymph node revealed cell-specific staining that immunohistochemistry analyses. Since sensitive can be properly preserved tissues, it allows retrospective analyses archival material.