| Objective:Research on potential of the DATS reverse multi drug resistance of breast cancer stem cell, then compared the cells treatment before and after the sensitivity of chemotherapeutic drugs. Research the molecular mechanism of multi drug resistance of DATS reversal,Whether because of abnormal inhibiting activation of NF- kappa B, and then it decreased P-gp expression, thus reversing the multidrug resistance of breast cancer stem cells.Methods:First of all, the detection of breast cancer stem cells in the amount of two kinds of MCF-7 and MCF-7/ADM cells by adherent culture, balloon culture and flow cytometer, then using the balloon culturing methods to collect breast cancer stem cells of MCF-7/ADM. After that, using MTT method to detect the DATS toxicity to the MCF-7/ADM balloon cells(breast cancer stem cells) and adherent cells(breast cancer cell), and the role of flow cytometry was used to detect ADM on cell toxicity which before and after the use of DATS to reverse drug resistance; then, regulating effect on the expression of P-gp and NF- κB Western Blotting for detection of DATS, at last, all the statistics using the SPSS17.0 statistical software.Results:(1)MCF-7 cells and MCF-7/ADM cells began to form a balloon in training third days, tenth days balloon formation rate were 1.13±0.55% and 10.16±0.54%, Further on the use of flow cytometry, two kinds of CD44+CD24- cell lines in the proportion were 2.83±1.38% and 63.94±2.31%. MCF-7/ADM cells in breast cancer stem cell content is higher, the difference was statistically significant. Both the detection results of si milar culture method.(2)In the DATS treatment of different concentrations of 24 h, MCF-7/ADM in adherent cells with balloon cell survival rate showed no significant difference in 10, 20μmol/L,When the concentration of DATS was 40, 60, 80 μmol/L, MCF-7/ADM balloon cell survival rate was significantly higher than that of the adherent cells, and the difference has statistical significance. The concentration of DATS in 60 μmol/L, two groups of cell apoptosis.(3)When the DATS concentration is 10 mol/L with ADM, the adherent cells and balloon cells’ s apoptosis rates(41.07 ± 0.29%, 10.30 ± 0.79%) much higher than using ADM alone group(25.46 ± 0.58%, 5.42 ± 0.31%), and the difference is statistically significant(P<0.05, n=3). After removed the drugs about 24 h,usings ADM alone groups’ apoptosis rate decreased, and DATS with ADM groups’ apoptosis rate rose slightly, the difference was significant, after joining DATS, the relative increase balloon cell sensitivity to ADM(4)MCF-7/ADM balloon cells than adherent cells’ P-gp and NF- kappa B expression is more obvious; After joining the DATS, the two kinds of cells’ P-gp and NF- kappa B was inhibited, the difference was statistically significant(P<0.05, n=3); The use of ADM and DATS after 24 h, two groups of cells P-gp and NF- kappa B expression was more inhibited.(5)Different concentrations of PDTC(NF- kappa B inhibitor) treatment of two kinds of cells by 24 h, with Western Blotting detection. DATS interacts with 40μmol/L PDTC, two groups of cells’ P-gp expression was inhibited in a dose-dependent manner, the difference was significant. Cells were 40μmol/L PDTC+DATS and 0μmol/LPDTC+DATS after 24 h treatment, the former than the latter’s P-gp expression was inhibited more significantly(P<0.05);Two groups of cells were added 40μmol/L PDTC with DATS, after 0h, 12 h, 24 h, 48 h, P-gp has obvious inhibition and depends on the concentration, the difference was significant(P<0.05).Conclusions:(1)MCF-7/ADM cell suspension culture can get breast cancer stem cells(2)DATS can reverse the breast cancer stem cells to adriamycin resistant effect.(3)P-gp and NF- kappa B high expression relate to breast cancer stem cells resistant.(4)DATS interference NF- kappa B, which can inhibit the activation of P-gp, and reverse the breast cancer stem cell resistance to chemotherapy. |
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