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Objective: Breast cancer is a highly heterogeneous common malignant tumor incidence increased year by year. Triple negative breast cancer (triple-negative breast cancer, TNBC) refers to the tumor cell surface lack of estrogen receptors (estrogen receptor, ER), progesterone receptor (progesterone receptor, PR) and human epidermal growth factor receptor 2 (human epidermal growth factor receptor-2, HER2) is a class of breast cancer, with postoperative recurrence, distant metastasis and poor prognosis easily the characteristics of endocrine therapy and targeted therapy for HER2, it still lacks a satisfactory comprehensive treatment program. Proposed in recent years cancer stem cells (cancer stem cell CSC) theory that cancer stem cells are the root causes of cancer, so TNBC various chemotherapeutic drugs on cancer stem cells are critically important, and its treatment may ultimately determine the success or failure of TNBC. At present, most studies show that CD44 CD24 -/low phenotype of breast cancer stem cells subsets. This study was six kinds of advanced breast cancer chemotherapy drugs commonly used triple-negative breast cancer cell line MDA-MB-231's role, and to observe these drugs on breast cancer stem cell subsets (CD44 CD24 -/low cells) content changes, the simultaneous detection of 10 cases of advanced metastatic triple-negative breast cancer patients before and after chemotherapy in peripheral blood CD44 CD24 -/low subsets levels in order for the clinical treatment of TNBC provide the basis for drug selection. Methods: a cultured human triple negative breast cancer cell line MDA-MB-231, use six kinds of advanced breast cancer chemotherapy drugs commonly used are: paclitaxel (PTX), docetaxel (DOX), 5 - fluorouracil (5 - FU), epirubicin (EPI), vinorelbine (NVB), gemcitabine (GEM). Application tetrazolium blue (tetrzolium-based colorimetric assay, MTT) method for the determination of various drugs drug concentration 0.2,1,5 times peak plasma concentrations (PPC), respectively, the role of 24h, 48h and 72h on MDA-MB -231 cell proliferation inhibition rate. 2 cells were randomly divided into control group (cultured under normal conditions MDA-MB-231 cell line) and six kinds of chemotherapy drugs in rats groups were also set up 10 cell culture flasks. Six kinds of chemotherapy drugs are a double PPC as a working concentration, were acting on the MDA-MB-231, 48 hours later, the cells were observed morphological changes, and five bottles of cells in each group was detected by flow cytometry CD44 CD24 -/low cell ratio; remaining five bottles of cells in the drug 48 hours after the removal of drug effects, adding a new medium, under normal culture conditions, cells cultured to recovery to bottle cells were observed in the number of adherent cells accounted for 80% of the bottom area of the standard, and then each group was detected by flow cytometry in CD44 CD24 -/low < / sup> cell ratio. 3 flow cytometry 10 cases of advanced metastatic triple-negative breast cancer patients before chemotherapy and after chemotherapy peripheral blood CD44 CD24 -/low cell levels, another collection 10 healthy volunteers blood samples as normal controls for comparison. Results: 1 MTT analysis showed that compared with control group, PTX, DOX, EPI ,5-FU, NVB, GEM in 0.2,1,5 times PPC 24,48,72 hours, the MDA-MB-231 significantly inhibited cell proliferation (p lt; 0.05), and with the increase of drug concentration, the inhibition rate also increases. 1 times the PPC for 48 hours, the MDA-MB-231 inhibition with PTX most obvious. 2 Flow cytometry results showed that: the control group CD44 CD24 -/low cell content: 86.46% ± 4.72%. PPC in a fold of each drug after 48h of CD44 CD24 -/low cell subsets contents were: NVB (91.34% ± 4.17%), GEM (85.43% ± 2.76%), DOX (74.78% ± 3.98%), PTX (71.67% ± 1.87%), EPI (66.38% ± 3.06%) ,5-FU (42.41% ± 5.43%). Statistics showed that the role and although it causes cells NVB content increased, compared with the control group, but the difference was not statistically significant (P gt; 0.05), GEM role, not the cell contents change significantly compared with the control group, the difference was not statistically significant (P gt; 0.05), the remaining contents of the cells in each group were reduced compared with the control group, the difference was statistically significant (p lt; 0.05), in which the effect of 5-FU after reducing the most obvious. Termination of drug effects, under normal conditions, cells cultured to recovery, flow cytometry in each group CD44 CD24 -/low cell contents were: NVB (86.46% ± 2.58%), GEM (85.97% ± 3.80%), DOX (75.23% ± 2.91%), PTX (69.21% ± 5.55%). Statistics showed NVB group CD44 CD24 -/low cell levels similar to the normal control, the difference was not statistically significant (P gt; 0.05), GEM group were content with the control group no statistically significant difference compared (P gt; 0.05), DOX and PTX group cell levels below normal, compared with the control group, the difference was statistically significant (p lt; 0.05). EPI and 5-FU group of cells did not resume normal growth (repeated training three times). 310 patients after two cycles of chemotherapy efficacy evaluation were stable disease (SD, in varying degrees of tumor shrink). 10 cases of advanced metastatic triple-negative breast cancer patients before chemotherapy and peripheral blood after chemotherapy CD44 CD24 -/low cell contents were: 54 ~ 793 / 100ul and 13 ~ 198 / 100ul, median, respectively: 160 / 100ul and 48 / 100ul. Peripheral blood of 10 healthy volunteers CD44 CD24 -/low cell content of 13 to 41 / 100ul, the median is 28 / 100ul. Statistics showed that in patients with metastatic triple-negative breast cancer in peripheral blood CD44 CD24 -/low cells were higher than those in healthy volunteers (p lt; 0.05). Peripheral blood of breast cancer patients after chemotherapy CD44 CD24 -/low cell levels lower than before chemotherapy (p lt; 0.05). Conclusion: a six kinds of chemotherapy drugs in 0.2,1,5 times PPC 24,48,72 hours, the MDA-MB-231 cells were significantly inhibited, with the increase of drug concentration, the inhibition rate increases. 1 times the PPC for 48 hours, the MDA-MB-231 inhibition with PTX most obvious. 2 triple negative breast cancer cell line MDA-MB-231 in CD44 CD24 -/low cell subsets of breast cancer stem cells subpopulation is not for all chemotherapy drugs induce resistance , chemotherapy drugs PTX, DOX, EPI ,5-FU that the content can be reduced to the most obvious effect of 5-FU. 3 advanced metastatic triple-negative breast cancer patients blood CD44 CD24 -/low subsets of breast cancer stem cells were higher than in healthy volunteers. Peripheral blood of breast cancer patients chemotherapy can CD44 CD24 -/low cells decreased. Breast cancer stem cells in the blood content and related subsets in patients with metastatic breast cancer tumor burden has some relevance, could become a very valuable clinical effect judgment index.
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