Detection of circulating breast cancer cells with multiple-marker RT-PCR assay.
RT-PCR assay for multiple markers has been shown to increase the detection rate of circulating tumor cells. This assay targeted against cytokeratin 19 (CK19), cytokeratin 20 (CK20) and beta-subunit of human chorionic gonadotropin (beta-hCG) was used to detect circulating breast cancer cells. 5 ml of peripheral blood was drawn before any surgical procedures from 72 breast cancer patients and 30 cases with benign breast disease. Total RNA was extracted from peripheral blood mononuclear cells, reverse-transcripted and amplified. For the benign cases, 10% (3/30) were positive for CK19 and all were negative for CK20 and beta-hCG, whereas 9.72% (7/72), 2.78% (2/72) and 12.5% (9/72) of the malignant cases were positive for CK19, CK20 and beta-hCG respectively. The detection rate of circulating breast cancer cells was unchanged when CK19 was combined with CK20, but it increased to 18.1% (13/72) when the marker was combined with beta-hCG. A significant difference was observed for beta-hCG between benign cases and affected patients with stage II, III and IV disease (p = 0.026). In conclusion, positive RT-PCR signals in blood samples of affected patients correlated with stage, in particular for beta-hCG. CK19/beta-hCG was a promising marker combination.
Hu
X C
XC
Department of Surgery, University of Hong Kong Medical Centre, Queen Mary Hospital, Pokfulam, Hong Kong.
Chow
L W
LW
D000328:Adult; D000368:Aged; D001943:Breast Neoplasms / Q000175:diagnosis* / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D018997:Chorionic Gonadotropin, beta Subunit, Human / Q000096:biosynthesis* / Q000235:genetics; D005260:Female; D006801:Humans; D007381:Intermediate Filament Proteins / Q000096:biosynthesis* / Q000235:genetics; D053554:Keratin-20; D007633:Keratins / Q000096:biosynthesis* / Q000235:genetics; D008875:Middle Aged; D009360:Neoplastic Cells, Circulating / Q000378:metabolism*; D011379:Prognosis; D012333:RNA, Messenger / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction
D018997:Chorionic Gonadotropin, beta Subunit, Human; D007381:Intermediate Filament Proteins; C507499:KRT20 protein, human; D053554:Keratin-20; D012333:RNA, Messenger; D007633:Keratins