Detection of disseminated tumor cells in bone marrow of gastric cancer using magnetic activated cell sorting and fluorescent activated cell sorting.
OBJECTIVE
Magnetic activated cell sorting (MACS) and fluorescent activated cell sorting (FACS) were employed to enrich and detect the gastric cancer cells from a cell line in a model system, and to enrich and detect disseminated tumor cells (DTCs) from bone marrow (BM) of patients with gastric cancer.


METHODS
Fifteen patients with benign gastric lesions and 35 patients with gastric cancer who received curative operations between December 2002 and June 2003 were selected. Mononuclear cells were separated from their BM. Cells from cell line OCUM-2M were seeded with 10-grade ratio into mononuclear cells from patients with benign gastric lesion. After labeling by MACS minibeads conjugated with cytokeratin (CK) 7/8 antibodies, anti-CK-fluorescein isothiocyanate (FITC), and anti-CD45-perdinin chlorophyll protein (PerCP), the samples were enriched twice using an MS+/RS+ positive separation column. The FACS analysis was conducted on these samples before and after MACS enrichment. The results were analyzed using clinopathological parameters.


RESULTS
Disseminated tumor cells were detected in the BM of 25 (71.43%) patients with gastric cancer. The frequencies of DTCs were 1.38 x 10(-8)-2.40 x 10(-5), 2.19 x 10(-7)-3.70 x 10(-5), 4.01 x 10(-6)-8.57 x 10(-5) in patients with well, moderately, and poorly differentiated carcinoma, respectively (P = 0.026). Disseminated tumor cells in BM had close correlation with tumor tumor-node-metastasis (TNM) stage (P = 0.034) and cancer-free survival (P = 0.035).


CONCLUSIONS
Disseminated tumor cells are very common in the BM of gastric cancer patients. Poor histological differentiation and more advanced TNM stage have more DTCs in the BM of gastric cancer patients. Patients with DTCs tend to have a poor prognosis.
Wang
Gui-Ying
GY
Department of Surgery, Fourth Hospital of Hebei Medical University, Shijiazhuang, Hebei, China.
Li
Yong
Y
Yu
Yue-Ming
YM
Yu
Bin
B
Zhang
Zhen-Ya
ZY
Liu
Yu
Y
Wang
Shi-Jie
SJ
D000328:Adult; D000368:Aged; D001854:Bone Marrow Cells / Q000737:chemistry / Q000473:pathology*; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D002469:Cell Separation / Q000379:methods*; D005260:Female; D005434:Flow Cytometry; D005743:Gastrectomy; D006801:Humans; D018189:Immunomagnetic Separation; D053208:Kaplan-Meier Estimate; D053552:Keratin-7 / Q000032:analysis; D053533:Keratin-8 / Q000032:analysis; D017493:Leukocyte Common Antigens / Q000032:analysis; D008297:Male; D008875:Middle Aged; D009362:Neoplasm Metastasis; D009367:Neoplasm Staging; D013274:Stomach Neoplasms / Q000737:chemistry / Q000401:mortality / Q000473:pathology* / Q000601:surgery; D013997:Time Factors; D016896:Treatment Outcome
C507497:KRT7 protein, human; C426203:KRT8 protein, human; D053552:Keratin-7; D053533:Keratin-8; D017493:Leukocyte Common Antigens; C516769:PTPRC protein, human