As shown in Figure 5, fragmented DNA was barely detectable. However, substantial amounts of low molecular weight DNA were present. indicating that either a small subset of cells had undergone internucleosomal DNA di gestion or that only a fraction of each cells DNA had be come fragmented. Although DNA fragmentation activator Ivacaftor has been seen in many cell types and Inhibitors,Modulators,Libraries is generally considered the bio chemical hallmark of apoptosis, it may be delayed, partial or absent in some cell types or experimental conditions. Therefore, it seems that treatment of MCF 7 and MDA MB 231 cells with these doses not leads to significant DNA fragmentation. Previous studies show also that treatment of epithelial cancer cell lines with a specific DNA damaging agent will produce high molecular weight DNA fragmentation in the absence of nucleosomal laddering.
In Addition, some apoptosis studies fail to exhibit the DNA fragmentation pattern in the mammary carcinoma cells. Levels of bax and bcl 2 mRNA expression To further investigate the apoptotic action of these two agents, we used Inhibitors,Modulators,Libraries quantitative real time PCR to study the influence of them on bcl 2 and bax mRNA expression. In many human cancers, the anti apoptotic bcl 2 pro teins are overexpressed, or the pro apoptotic proteins like bax, have reduced expression. This results in re sistance to a wide variety of cell death stimuli including chemotherapeutic drugs. Inhibitors,Modulators,Libraries Results of real time quantitative PCR appeared to show down regulation Inhibitors,Modulators,Libraries of bcl 2 and upregulation of bax expres sion at 48 hours treatment.
Expression of bcl 2 and bax was targets for TAM and tranilast as a sin gle or combination and after 48 h exposure, a significant reduction of bcl Inhibitors,Modulators,Libraries 2 and induction of bax mRNA expression was observed. Bax to bcl 2 mRNA ratio was determined for MCF 7 cells 3. 4 in TAM treatment, 3. 0 in tranilast treatment and 8. 4 in combined group and for MDA MB 231 cells 1. 7 in TAM sellectchem treatment, 2. 2 in tranilast treatment and 3. 8 in combination. Hence, the ratio of pro apoptotic to the anti apoptotic was altered in favor of apoptosis. Thus, the results suggest that an up regulation of bax and the corresponding down regulation of bcl 2 mRNAs observed in this study may be one of the critical mechanisms through which TAM and/or tranilast induces apoptosis in breast cancer cells. Effects of TAM and/or tranilast treatment on mRNA level of TGF B ligands and receptors in breast cancer cells Exposure of cell cultures to TAM and tranilast either alone or in combination for 48 h decreased expression of TGF B1, B2, B3 and TBRI, BRII mRNA. TGF B1 mRNA levels were high but 48 h after TAM, tranilast or combined treatment they were diminished approxi mately a 30%, 70% and 92% in MCF 7 cells and 15%, 40% and 60% in MDA MB 231 cells.