Evaluation of circulating tumour cells expressing CD44 variants in the blood of gastric cancer patients by flow cytometry.
The occurrence of circulating tumour cells in the blood of 51 patients with gastric cancer (stages I-IV) was studied using flow cytometry, cell sorting, immunohistochemistry and reverse transcription polymerase chain reaction (RT-PCR). The lysed whole blood samples were stained with monoclonal antibodies against common leukocyte antigen (CD45), epithelial membrane antigen (EMA), tumour associated glycoprotein 72kD (TAG72), CD44 variants (v5 and v6) and analysed by flow cytometry within ungated or CD45-gated populations. The frequency of detection of TAG72+, CD44v5+ and v6+ cells within CD45- gate was considerably increased in comparison to the ungated population. Furthermore, the presence of tumour cells was directly demonstrated by immunostaining for cytokeratin 18 of sorted CD45- population. The presence of CD44v5+, v6- cells and CD44v-mRNA in the blood was compared to their expression in the primary tumour. The occurrence of circulating CD44v+ cells was associated with their presence in the primary tumour and CD44v-mRNA in the blood. The method described may provide a sensitive tumour marker-independent tool for detection of circulating tumour cells in cancer patients.
Pituch-Noworolska
A
A
Department of Clinical Immunology, Polish-American Institute of Paediatrics, Cracow, Poland.
Wieckiewicz
J
J
Krzeszowiak
A
A
Stachura
J
J
Ruggiero
I
I
Gawlicka
M
M
Szczepanik
A
A
Karcz
D
D
Popiela
T
T
Zembala
M
M
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000951:Antigens, Neoplasm / Q000097:blood / Q000276:immunology; D014408:Biomarkers, Tumor / Q000097:blood*; D002469:Cell Separation; D005260:Female; D005434:Flow Cytometry / Q000379:methods; D006023:Glycoproteins / Q000097:blood / Q000276:immunology; D006801:Humans; D018960:Hyaluronan Receptors / Q000097:blood* / Q000276:immunology; D017493:Leukocyte Common Antigens / Q000097:blood / Q000276:immunology; D007963:Leukocytes, Mononuclear / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D018396:Mucin-1 / Q000097:blood / Q000276:immunology; D009360:Neoplastic Cells, Circulating / Q000276:immunology*; D013274:Stomach Neoplasms / Q000097:blood* / Q000276:immunology / Q000378:metabolism / Q000473:pathology
D000951:Antigens, Neoplasm; D014408:Biomarkers, Tumor; D006023:Glycoproteins; D018960:Hyaluronan Receptors; D018396:Mucin-1; C047052:tumor-associated antigen 72; D017493:Leukocyte Common Antigens