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ABSTRACT: Circulating tumor stem cells (CTSC), a subpopulation of circulating tumor cells (CTC), may lead to recurrent diseases. The aim of this study was to detect CTC (CD45(-)EpCAM(+)) and CTSC (CD45(-)EpCAM(+)CD44(+)CD24(-)) of breast cancer (BC) patients, as well as to explore their clinical relevance. CTC and CTSC in peripheral blood (PB) of 45 female BC patients were detected by using flow cytometry (FCM). SKBR-3 cells were mixed with MNC of four healthy volunteers at different ratios in order to evaluate the sensitivity of FCM. Real-time quantitative polymerase chain reaction (QRT-PCR) was conducted and compared with FCM. The expression of EPCAM between CTC < 50 and CTC ≥ 50 groups (19.98 ± 23.93 versus 29.46 ± 29.27 × 10(-5)), and the expression of CD44 between CTSC negative and positive groups (0.85 ± 0.91 versus 0.81 ± 0.75) were statistically the same. FCM had higher specificity than QRT-PCR. Statistical differences were obtained between CTC < 50 and CTC ≥ 50 groups among different TNM stages, histology stages, estrogen receptor (ER) status and progesterone receptor (PR) status (P < 0.05). Statistical differences between CTSC negative and positive groups within different TNM stages and regional lymph node metastasis (RLNM) status (P < 0.05) were also obtained. Moreover, the percentage of CTC on CD45 negative cells (CD45(-)C) among different clinical pathology was statistically different, P = 0.000. Additionally, the percentage of CTSC on CD45(-)C with TNM stage was rising (0: 0.00 ± 0.00‰, I: 0.03 ± 0.05‰, II: 0.06 ± 0.14‰, III: 0.10 ± 0.09‰, IV: 0.29 ± 0.35‰, P = 0.034). Statistical difference in the percentage of CTSC on CD45(-)C among different RLNM status (P = 0.001) was also obtained. FCM to detect CTC and CTSC may be used to diagnose disease at early stage, to guide clinical therapy or to predict prognosis.
Tumor Biology 04/2012; 33(2):561-9. · 1.94 Impact Factor
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ABSTRACT: We wanted to demonstrate the value of multiparameter flow cytometry in detecting human tumor cells of breast cancer (BC) (SKBR-3) in normal peripheral blood. In addition, we investigated a cluster of patients to compare the overall survival (OS) between advanced BC patients [circulating tumor cells (CTCs) >or=5 group] and limited BC patients (CTCs <5 group). SKBR-3 human BC cells were serially diluted in normal whole blood to demonstrate the sensitivity of multiparameter flow cytometry for detecting CTCs, and we also compared the specificity with reverse transcriptase polymerase chain reaction (RT-PCR) method. On the other hand, we detected CTCs among 45 patients by multiparameter flow cytometry. OS was calculated by the Kaplan-Meier product limit method, and compared it between CTCs <5 and CTCs >or=5 groups with the log-rank test. Cox regression models were fitted to determine the associated factors on survival. Human BC cells (SKBR-3) could be differentiated from normal blood based on the multiple light scatter and cell surface marker expression by multiparameter flow cytometry. The method was found to have a sensitivity limit of 10(-5) and was effective for detecting human BC cells in vivo. It also found that this method had a higher specificity compared with RT-PCR. For the retrospective study, the median OS was 95 weeks and 65.5 weeks (P < 0.05, 2-tailed) for patients with CTCs <5 and CTCs >or=5, respectively. Kaplan-Meier was used to analyze the patients' survival with Log Rank P = 0.004 and Breslow P = 0.003, which showed that these two groups had statistically significant difference. Cox regression analysis was performed, and we found CTCslevels, metastasis and age (P < 0.05) were three relative factors for patients' survival. Multiparameter flow cytometry can detect CTCs effectively and has the potential to be a valuable tool for prognosis assessment among BC patients in clinical situations in China.
Cytometry Part A 02/2010; 77(3):213-9. · 3.73 Impact Factor
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ABSTRACT: To investigate the characteristics of T lymphocytic subsets in chronic tonsillitis patients for evaluation of their clinical implication.
Fresh peripheral blood samples were obtained from 54 chronic tonsillitis patients and 52 healthy counterparts. CD4+ and CD8+ T lymphocyte subsets including the naive (CD45RA+), memory (CD45RO+), functional (CD28+), activated (HLA-DR+, CD25+) and apoptosis (CD95+) T lymphocytes, were analyzed by flow cytometry, respectively. The clinical data such as serum Cystatin C con centration, ASO and ESR were simultaneously recorded from each chronic tonsillitis patient.
The CD4+ T cells, the rate of CD4+/CD8+ and CD4+ CD45RA+ /CD4+ CD45RO+, the CD4+ CD45RA+ T cells and the CD4+ CD25+ T cells in chronic tonsillitis were significantly lower than those of the control group (P < 0.05) while an obviously increasing percentage of memory (CD45RO+) and apoptosis (CD95+) T lymphocytes in chronic tonsillitis patients, and there were significant differences between the patients with chronic tonsillitis and the healthy volunteers (P < 0.05). Furthermore, in the chronic tonsillitis patients, Serum Cystatin C level was negatively correlated with CD4+ CD45RA+ T(P < 0.05), and not significantly correlated with other T lymphocyte subtypes (P > 0.05).
Immune disorder is present in the peripheral blood of chronic tonsillitis patients. Our data may provide valuable information for evaluation of disease progression of chronic tonsillitis patients.
Lin chuang er bi yan hou ke za zhi = Journal of clinical otorhinolaryngology 06/2009; 23(10):436-8, 442.
