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Fig. 5. Effect of PD98059 Parke-Davis (PD) on DFMO-induced suppression of invasiveness in the presence (A) or in the absence (B) of hepatocyte growth factor (HGF) (40 ng/mL) as a chemoattractant. MDA-MB-435 breast cancer cells were cultured in regular medium in the absence and in the presence of DFMO (1 mM) for 48 h before plating in the matrigel assay. PD treatment (50 |M was added 1 h before plating in the matrigel assay (time found to be sufficient to block extracellular regulated kinase phosphorylation) and continued for the duration of the experiment. Data are expressed as percent of control. Bars represent means ± standard error of the mean. The number of replicate experiments is five in (A) and four in (B). Both in the presence (A) and in the absence of HGF (B), treatment with PD significantly reversed the anti-invasive effect of DFMO (p < 0.05). (Reproduced from ref. 46 with permission of Springer Science and Business Media.)

Fig. 6. Effect of PD98059 Parke-Davis (PD) on DFMO-induced stimulation of throm-bospondin-1 (TSP-1) production. MDA-MB-435 breast cancer cells were cultured for 48 h in regular medium in the absence and in the presence of DFMO. The cells were then stepped down to serum-free OPTI-MEM containing the same treatments for 48 h. PD98059 was added 1 h before transition to serum-free media conditions and continued until the end of the experiment. The conditioned media were collected, processed, and subjected to Western analysis using an antibody against TSP-1 as described (46). (Reproduced from ref. 46 with permission of Springer Science and Business Media.)

Fig. 6. Effect of PD98059 Parke-Davis (PD) on DFMO-induced stimulation of throm-bospondin-1 (TSP-1) production. MDA-MB-435 breast cancer cells were cultured for 48 h in regular medium in the absence and in the presence of DFMO. The cells were then stepped down to serum-free OPTI-MEM containing the same treatments for 48 h. PD98059 was added 1 h before transition to serum-free media conditions and continued until the end of the experiment. The conditioned media were collected, processed, and subjected to Western analysis using an antibody against TSP-1 as described (46). (Reproduced from ref. 46 with permission of Springer Science and Business Media.)

increased p21 expression, reduced cdk-2 protein and activity, reduction in the phosphorylation of the retinoblastoma gene, and G1/S arrest. These results are in general agreement with similar observations reported in melanoma and intestinal epithelial cells (48,49).

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