Therefore, we investigated whether http://www.selleckchem.com/products/nutlin-3a.html doxorubicin has the same anti proliferative 17-AAG manufacturer effect in MDA MB 231, MCF 7, MCF 7/ DOX, and T 47D cell lines. The cells Inhibitors,Modulators,Libraries were treated with the indicated concentrations of doxorubicin for 72 h and cell proliferation was measured using the MTT assay. Doxorubicin Inhibitors,Modulators,Libraries inhibited moreover cell proliferation in a concentra tion dependent manner in MCF 7 and T47D cells, and to a lesser extent in MDA MB 231 cells. By contrast, doxorubicin mediated inhibition was significantly reduced in MCF 7/DOX cells. We next mea sured the growth of MCF 7 and MCF 7/DOX cells at lower doxorubicin Inhibitors,Modulators,Libraries concentrations and MCF 7/DOX cells were consistently resistant to doxorubicin.
We then tested whether the doxorubicin mediated growth inhibition was mediated by apoptosis.
After MCF 7 and MCF 7/DOX cells were treated with 1 uM Inhibitors,Modulators,Libraries Inhibitors,Modulators,Libraries doxorubicin for 48 h, terminal deoxynucleotidyl trans ferase mediated dUTP nick Inhibitors,Modulators,Libraries end labeling based fluores cence activated cell sorting analysis showed that doxorubicin Inhibitors,Modulators,Libraries did not induce apoptosis in MCF 7/DOX cells. however, doxorubicin did induce apoptosis in MCF 7 cells. We further confirmed the resistance of MCF 7/DOX cells to doxorubicin by Inhibitors,Modulators,Libraries Wes tern blot analysis. Induction of PARP cleavage in MCF 7 cells confirmed that doxorubicin induced apoptosis in these cells. However, PARP was not cleaved in MCF 7/DOX cells treated with doxorubi cin.
Acquisition of invasive and metastatic properties in MCF 7/DOX cells Intrinsic or acquired drug resistance Inhibitors,Modulators,Libraries results in disease recurrence and metastasis.
We analyzed changes in gene expression in doxorubicin resistant MCF 7/DOX cells using DNA array analysis.
Differentially Inhibitors,Modulators,Libraries expressed Inhibitors,Modulators,Libraries genes related with invasion are listed in Table 1. We next examined whether MCF 7/DOX cells acquired metastatic properties. First, we measured the Inhibitors,Modulators,Libraries enzymatic activities of MMP 9, MMP 2, and uPA in MCF 7 and MCF 7/DOX cells by gelatin and fibrino gen/plasminogen zymography. The enzymatic activities of MMP 2, MMP 9, and uPA were Inhibitors,Modulators,Libraries markedly higher in MCF 7/DOX cells than in non invasive MCF 7 cells. Increased levels of MMP 9 and MMP 2 expression have been correlated with an invasive pheno type of cancer cells.
Thus, we assessed the invasive ness of MCF 7 and MCF 7/DOX cells using in vitro invasion assays.
As expected, the MCF 7/DOX cells were significantly more invasive than MCF 7 Inhibitors,Modulators,Libraries cells.
selleck Tofacitinib To test the invasive activity selleck screening library of MCF 7/DOX cells in vivo, we developed a breast cancer model of lung metastasis. Three months after injecting MCF 7/ DOX cells through the tail veins Inhibitors,Modulators,Libraries of balb/c nude mice, the mice were killed, selleck chemicals and the total number of visible lung tumor nodules per mouse was quantified under a stereomicroscope. Lung tumor nodules were present in the mice injected with MCF 7/DOX cells, while no mouse injected with MCF 7 cells had lung metastases.