title: Patterns of circulating tumor cells identified by CEP8, CK and CD45 in pancreatic cancer.
abstract: To improve the identification for CTCs with weak or negative CK and diploid CTCs in pancreatic cancer, we combined immune-staining of CK, CD45, DAPI and fluorescence in situ hybridization with the centromere of chromosome 8 (CEP8) probe method. CTCs in 3.75 mL of blood were depleted for CD45 positive cells with anti-CD45 antibodies and identified by combining CK, CD45, DAPI and CEP8 in 61 cases including 22 pancreatic cancers, 3 borderline pancreatic solid pseudopapillary tumors, 6 pancreatic benign tumors, and 30 healthy individuals. We found that enriched cells could be classified into 5 patterns: CK+CD45-DAPI+CEP8=2 (2 hybridization signals), CK+CD45-DAPI+CEP8>2 (>2 hybridization signals), CK-CD45-DAPI+CEP8>2, CK-CD45-DAPI+CEP8=2, and CK+/-CD45+DAPI+CEP8=2 or >2. Among 22 pancreatic cancers, CK+CD45-DAPI+CEP8=2 and CK+CD45-DAPI+CEP8>2 patterns were identified in two cases, and CK-CD45- DAPI+CEP8>2 pattern was identified in 16 cases. CK-CD45-DAPI+CEP8=2 and CK+/-CD45+DAPI+CEP8=2 or >2 patterns were detected in pancreatic cancers, other pancreatic diseases and healthy individuals. Among the five patterns, CK+CD45-DAPI+CEP8=2, CK+CD45-DAPI+CEP8>2 and CK-CD45-DAPI+CEP8>2 were considered as CTCs, while CK-CD45-DAPI+CEP8=2 and CK+/-CD45+DAPI+CEP8=2 or >2 were considered as indeterminate cells. When the cutoff value was set as 2 cells/3.75 mL based on ROC curve, the sensitivity and specificity in the diagnosis of pancreatic cancer was 68.18 and 94.87%, respectively. Dynamically monitoring CTCs changes prior to and after surgery in pancreatic patients revealed that CTCs count decreased in 3 days after surgery, but increased in 10 days after surgery in most patients. During our one and a half year follow-up, CTCs positive patients showed metastasis and worse survival rate.
authors.affiliation: Department of Clinical Laboratory, Peking Union Medical College Hospital, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing, China.
authors.forename: Yujuan
authors.initials: Y
authors.lastname: Zhang
authors.forename: Fei
authors.initials: F
authors.lastname: Wang
authors.forename: Ning
authors.initials: N
authors.lastname: Ning
authors.forename: Qian
authors.initials: Q
authors.lastname: Chen
authors.forename: Zhuo
authors.initials: Z
authors.lastname: Yang
authors.forename: Ye
authors.initials: Y
authors.lastname: Guo
authors.forename: Danfei
authors.initials: D
authors.lastname: Xu
authors.forename: Donghong
authors.initials: D
authors.lastname: Zhang
authors.forename: Ting
authors.initials: T
authors.lastname: Zhan
authors.forename: Wei
authors.initials: W
authors.lastname: Cui
mesh_terms: D000368:Aged; D014408:Biomarkers, Tumor / Q000032:analysis*; D021441:Carcinoma, Pancreatic Ductal / Q000378:metabolism / Q000401:mortality / Q000556:secondary*; D016022:Case-Control Studies; D002503:Centromere / Q000235:genetics*; D002898:Chromosomes, Human, Pair 8 / Q000235:genetics; D005260:Female; D005500:Follow-Up Studies; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D007633:Keratins / Q000378:metabolism*; D017493:Leukocyte Common Antigens / Q000378:metabolism*; D008207:Lymphatic Metastasis; D008297:Male; D009364:Neoplasm Recurrence, Local / Q000378:metabolism / Q000401:mortality / Q000473:pathology*; D009367:Neoplasm Staging; D009360:Neoplastic Cells, Circulating / Q000473:pathology*; D010190:Pancreatic Neoplasms / Q000378:metabolism / Q000401:mortality / Q000473:pathology*; D011379:Prognosis; D015996:Survival Rate
chemical_list: D014408:Biomarkers, Tumor; D007633:Keratins; D017493:Leukocyte Common Antigens; C516769:PTPRC protein, human
keywords: CK; FISH; circulating tumor cells; pancreatic cancer; pattern