Selective expression of a splice variant of decay-accelerating factor in c-erbB-2-positive mammary carcinoma cells showing increased transendothelial invasiveness.
By differential-display-PCR a subclone of the SK-BR-3 cell line with high in vitro transendothelial invasiveness was identified to express increased levels of a new alternative splice variant of decay-accelerating factor (DAF). DAF seems to play an important role in some malignant tumours since on the one hand the expression of complement inhibitors on the surface of tumour cells prevents the accumulation of complement factors and in consequence cell lysis. On the other hand, DAF has been identified as a ligand for the CD97 surface receptor which induces cell migration. Immunofluorescence procedures, Western blot analyses, and cDNA clone sequencing were employed to confirm the expression of DAF restricted to invasive tumour cells. Using a radioactive RNA-in situ hybridisation on freshly frozen tissue microarrays and RT-PCR on native tumour tissue, the expression of alternative spliced DAF mRNA was demonstrated in invasive breast cancer. Due to the fact that it could thereby not be detected in normal mammary tissues, it has to be confirmed in larger studies that the DAF splice variant might be a specific tumour marker for invasive breast cancer.
Brandt
Burkhard
B
Institute of Clinical Chemistry and Laboratory Medicine, University of Muenster, Germany. brandt@uni-muenster.de
Mikesch
Jan-Hendrik
JH
Simon
Ronald
R
Rötger
Antje
A
Kemming
Dirk
D
Schier
Katrin
K
Sauter
Guido
G
Bürger
Horst
H
D017398:Alternative Splicing; D015703:Antigens, CD; D001483:Base Sequence; D014408:Biomarkers, Tumor; D015153:Blotting, Western; D001943:Breast Neoplasms / Q000378:metabolism*; D018958:CD55 Antigens / Q000096:biosynthesis* / Q000737:chemistry*; D002465:Cell Movement; D003001:Cloning, Molecular; D003169:Complement Inactivator Proteins / Q000737:chemistry; D003165:Complement System Proteins / Q000378:metabolism; D018076:DNA, Complementary / Q000378:metabolism; D004587:Electrophoresis, Agar Gel; D004730:Endothelium, Vascular / Q000166:cytology; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D008024:Ligands; D008562:Membrane Glycoproteins / Q000096:biosynthesis; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D009361:Neoplasm Invasiveness; D020411:Oligonucleotide Array Sequence Analysis; D016133:Polymerase Chain Reaction; D012313:RNA / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000096:biosynthesis* / Q000378:metabolism; D043562:Receptors, G-Protein-Coupled; D020133:Reverse Transcriptase Polymerase Chain Reaction
D015703:Antigens, CD; D014408:Biomarkers, Tumor; D018958:CD55 Antigens; D003169:Complement Inactivator Proteins; D003165:Complement System Proteins; D018076:DNA, Complementary; D008024:Ligands; D008562:Membrane Glycoproteins; D012313:RNA; D012333:RNA, Messenger; D018719:Erb-b2 Receptor Tyrosine Kinases; D043562:Receptors, G-Protein-Coupled; C095004:ADGRE5 protein, human