changes in protein profiles in bortezomib applied multiple myeloma ...
changes in protein profiles in bortezomib applied multiple myeloma ...
changes in protein profiles in bortezomib applied multiple myeloma ...
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4.2. Changes <strong>in</strong> Caspase-3 Enzyme Activity and Loss of Mithocondrial<br />
Membrane Potantial) MMP<br />
To evaluate the apoptotic effects of Bortezomib on U-266 cells, Caspase-3<br />
enzyme activity and mitochondrial membrane potential were two important apoptotic<br />
<strong>in</strong>dicators. Treatment of U-266 cells for 72 h with 1, 10 and 20 nM Bortezomib resulted<br />
<strong>in</strong> 1.06, 1.13 and 1.17-fold <strong>in</strong>creases, respectively, <strong>in</strong> caspase-3 activity as compared to<br />
untreated controls (Figure 4.2).<br />
Changes <strong>in</strong> Caspase-3 Enzyme<br />
Aktivity<br />
200<br />
150<br />
100<br />
50<br />
0<br />
100<br />
Caspase-3 Enzyme Aktivity<br />
106 113 117<br />
C 1 10 20<br />
Bortezomib (nM,72h)<br />
Figure 4.2. Changes <strong>in</strong> Caspase-3 Enzyme Activity<br />
We also assessed the loss of mitochondrial membrane potential <strong>in</strong> order to<br />
confirm the caspase-3 enzyme activity results and exam<strong>in</strong>e the roles of mitochondria <strong>in</strong><br />
Bortezomib-<strong>in</strong>duced apoptosis. Treatment of U-266 cells for 72 h with 1, 10 and 20 nM<br />
Bortezomib caused a significant loss of MMP (about 1.34, 1.85 and 2.14-fold,<br />
respectively, when compared with untreated controls), as measured by <strong>in</strong>creased<br />
accumulation of cytoplasmic mitochondrial JC-1 (Figure 4.3).<br />
U-266<br />
69