| Objective: This study analysis the relationship of circulating tumorcell(CTC) and tumor stem cells(TSC) in the breast cancer with clinical stages,histopathological index and prognosis, and explore the clinical value of usingflow cytometry to detect the breast cancer patients' peripheral bloodcirculating tumor cells and tumor stem cells.Methods:1Extracted peripheral blood10ml from60cases of primary breast cancerpatients who were treated in the fourth hospital breast center of HE BEImedical university from August2011to January2012, prepared theperipheral blood mononuclear cell(PBMC) by lymphocyte separation liquid,while extracted peripheral blood10ml from15cases of healthy volunteers ascontrol group.2A single nuclear peripheral blood cells were divided test tube and negativecontrol tube. The test tubes were respectively marked with cytokeratin19(CK19) mouse anti-human monoclonal antibody, CD44mouse anti-humanmonoclonal antibodies, CD24mouse anti-human monoclonal antibodies.The control tubers were respectively added isotype control antibodies ofCK19, CD44andCD24antibodies, the operation was completed In the sameconditions.3Peripheral blood mononuclear cells were stained with antibodies andanalyzed by flow cytometry, detected CK19+and CK19+/CD44+/CD24-/low proportion of cells, respectively, as the sign of peripheral bloodcirculating tumor cells and cancer stem cells.4The results were processes with SPSS13.0statistical software package,the experiment results applied X2inspection of statistical analysis, with P <0.05for differences with a statistical significance. Results:132cases were detected CTC in peripheral blood from60primary breastcancer patients, the positive rate of CTC was53.33%(32/60); In15cases ofhealthy volunteers, one case was detected CTC in peripheral blood, thepositive rate of CTC was6.67%(1/15), the differences of positive circulatingtumor cell had a statistics difference between the two groups (P=0.001).260patients with primary breast cancer, according to the clinical TNMstaging (Ⅰ, Ⅱ, Ⅲ and Ⅳ period) different, the positive rate of CTC were21.43%(3/14),54.17%(13/24),66.67%(12/18) and100.00%(4/4),respectively. There was significant difference in positive rate of CTC betweendifferent TNM stages (P=0.006). The positive rates of CTC in patients atpathologic lymph node stages (pN0, pN1-3) were34.62%(9/26),67.65%(23/34), respectively. There was significant difference in positive rate of CTCbetween different pN stages (P=0.018). According to the clinical primarytumor size (T1,T2,T3, T4) different, the positive rates of CTC in patientswere40.00%(8/20),61.11%(11/18),60.00%(12/20),50.00%(1/2),respectively. There was no significant difference in positive rate of CTCbetween different groups (P>0.05); The positive rates of CTC in ER negativepatients was66.67%(12/18), which was much higher than the ER positivepatients with the positive rate of47.62%(20/42); PR negative patients whosepositive rate of CTC was68.42%(13/19) were much higher than PR positivepatients, it was46.34%(19/41), but both were no statistical difference (P>0.05). The positive rate of CTC in patients with C-erbB2negative andC-erbB2positive was66.67%(8/12) and55.00%(22/40), but there was nostatistical difference (P>0.05); the positive rate of CTC in patients withLuminal, HER-2and Basle-like subtype was41.38%(12/29),80.00%(8/10)and76.92%(10/13) respectively. There was statistical difference in positiverate of CTC between different molecular subtypes (P=0.0240). The positiverate of CTC in patients with HER-2and Basle-like subtype was much higherthan patients with Luminal subtype, but there was no statistical differencebetween HER-2type and Luminal type (P>0.05); the positive rate of CTC in patients Basle-like subtype was significantly higher than patients withLuminal subtype (P=0.047).3In the32patients with the CTC positive,15cases were detectedcirculating tumor stem cells(CTSC), according to the clinical TNM staging(Ⅰ, Ⅱ, Ⅲ and Ⅳ period) different, the positive rate of circulating tumor stemcells were33.33%(1/3),38.46%(5/13),50.00%(6/12) and75.00%(3/4)respectively, along with the increase of the clinical stages of positive trend hadincreased, but there was no significant difference in positive rate of CTSCbetween different TNM stages (P>0.05). The positive rates of CTSC inpatients at pathologic lymph node stages (pN0, pN1-3) were55.56%(5/9),43.48%(10/23), there was no significant difference between different pNstages (P>0.05). According to the clinical primary tumor size (T1, T2, T3, T4)different, the positive rates of CTSC in patients were37.50%(3/8),45.45%(5/11),58.33%(7/12),0%(0/1) respectively, there was no significantdifference between different groups (P>0.05); the positive rates of CTSC inER negative patients was50.00%(6/12), which was much higher than the ERpositive patients with the positive rate of45.00%(9/20); the positive rate ofCTSC in patients with PR negative and PR positive was46.15%(6/13) and47.37%(9/19), but both were no statistical difference (P>0.05). The positiverate of CTSC in patients with C-erbB2negative and C-erbB2positive was45.45%(10/22) and62.50%(5/8), but there was no statistical difference (P>0.05); the positive rate of CTSC in patients with Luminal, HER-2andBasle-like subtype was35.71%(5/14),66.67%(4/6) and60.00%(6/10)respectively. The positive rate of CTSC in patients with HER-2and Basle-likesubtype was much higher than patients with Luminal subtype, but there wasno statistical difference between groups (P>0.05).Conclusions:1Using flow cytometry technology, circulating tumor cells and cancer stemcells can be detected from peripheral blood of primary breast cancer patients.2This research shows that the breast cancer patients' peripheral bloodcirculating tumor cells are relate to the bad breast cancer biology. The clinical stages the later, the higher pN staging, the positive rate of CTC in patients ishigher, and has a statistical significance. And it is no relevance with theprimary tumor size. Breast cancer patients' peripheral blood circulating tumorstem cells with the later clinical stages, the higher pN stages, and the higherthe positive, but there is no statistical significance.3The positive rate of CTC in patients is familiar related to ER, PR,C-erbB2molecular classification, The positive rates of CTC in ER negativepatients is much higher than the ER positive patients and it is also higher inPR negative patients and C-erbB2positive patients, but there is no statisticaldifference (P>0.05); the positive rate of CTC in patients with HER-2andBasle-like subtype is much higher than patients with Luminal subtype, butthere was no statistical difference between HER-2type and Luminal type (P>0.05); the positive rate of CTC in patients Basle-like subtype is significantlyhigher than patients with Luminal subtype (P=0.047).4The positive rate of CTSC in patients is familiar related to ER, PR,C-erbB2molecular classification, the positive rate of CTSC in ER negativepatients is much higher than the ER positive patients and it is higher in PRpositive patients and C-erbB2positive patients, but there is no statisticaldifference (P>0.05); The positive rate of CTSC in patients with HER-2andBasle-like subtype is much higher than patients with Luminal subtype, butthere is no statistical difference between groups (P>0.05). |
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