Identification of PECAM-1 in solid tumor cells and its potential involvement in tumor cell adhesion to endothelium.
PECAM-1 (CD31/EndoCAM) is an adhesion molecule in the immunoglobulin supergene family that is expressed on endothelial cells, platelets, and some hematopoietic lineage cells. In this paper, using several polyclonal and monoclonal antibodies against PECAM-1, we identified PECAM-1 molecules on human, rat, and murine solid tumor cell lines. Immunocytochemical labeling and flow cytometric analysis using either polyclonal, monoclonal, or Fab portion of the antibodies against PECAM-1 detected a distinct distribution on tumor cell surface. Immunoblotting revealed proteins ranging from 120 to 130 kDa in tumor cells derived from different species. Immunoprecipitation and subcellular fractionation studies indicated that PECAM-1 is constitutively expressed on the surface of human tumor cells (i.e. colon adenocarcinoma). The specificity of a major polyclonal anti-PECAM-1 used in the current study (i.e. SEW-3) was confirmed by the preabsorption studies. PECAM-1 molecules on tumor cells appear to bear terminal carbohydrate moieties (i.e. sialic acid residues) different from those on platelets, since neuraminidase treatment of tumor cells, unlike platelets, did not result in a mobility shift. Polymerase chain reaction (PCR) analysis of genomic DNA derived from tumor cell lines of different species revealed the presence of PECAM-1 gene in the genome. The mRNAs of PECAM-1 in tumor cells were detected by reverse transcription-PCR followed by Southern hybridization. Screening of more than 20 human, rat, and murine solid tumor cell lines indicated that PECAM-1 is widely expressed, although the level of expression varies considerably among different cell lines. The expression of PECAM-1 message in tumor cells was confirmed by Northern blotting. DNA sequencing of the PCR fragment revealed that human tumor cell PECAM-1 matches 100% to the human endothelial cell counterpart. Finally, it was demonstrated that tumor cell PECAM-1 is involved in mediating tumor cell adhesion to endothelium, as evidenced by the ability of anti-PECAM-1 antibodies to decrease the adhesion of unstimulated tumor cells to microvascular endothelial cells.
Tang
D G
DG
Department of Radiation Oncology, Wayne State University, Detroit, Michigan 48202.
Chen
Y Q
YQ
Newman
P J
PJ
Shi
L
L
Gao
X
X
Diglio
C A
CA
Honn
K V
KV
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies; D015214:Antigens, Differentiation, Myelomonocytic / Q000032:analysis* / Q000096:biosynthesis / Q000378:metabolism; D001483:Base Sequence; D002448:Cell Adhesion; D015815:Cell Adhesion Molecules / Q000032:analysis* / Q000378:metabolism; D002460:Cell Line; D002478:Cells, Cultured; D017931:DNA Primers; D004273:DNA, Neoplasm / Q000302:isolation & purification / Q000378:metabolism; D004730:Endothelium, Vascular / Q000502:physiology*; D005455:Fluorescent Antibody Technique; D006801:Humans; D007150:Immunohistochemistry; D051379:Mice; D008969:Molecular Sequence Data; D016376:Oligonucleotides, Antisense; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D016133:Polymerase Chain Reaction; D051381:Rats; D014407:Tumor Cells, Cultured / Q000166:cytology / Q000378:metabolism / Q000502:physiology*
D000906:Antibodies; D015214:Antigens, Differentiation, Myelomonocytic; D015815:Cell Adhesion Molecules; D017931:DNA Primers; D004273:DNA, Neoplasm; D016376:Oligonucleotides, Antisense; D019408:Platelet Endothelial Cell Adhesion Molecule-1