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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activity and loss of mitochondrial membrane potential (MMP). Treatment of U-266<br />
cells for 72 h with 20 nM Bortezomib caused a significant loss of MMP (about 2.14-<br />
fold), as measured by <strong>in</strong>creased accumulation of cytoplasmic mitochondrial form of JC-<br />
1 and also resulted <strong>in</strong> 1.17-fold <strong>in</strong>crease <strong>in</strong> caspase-3 activity.<br />
After the 2D PAGE analysis, so as to elucidate and compare prote<strong>in</strong> <strong>profiles</strong> of<br />
two groups at issue, two <strong>in</strong>dividual gel images were imported <strong>in</strong>to gel analysis software<br />
DECODON Delta2D Version 4.3. With the help of software, these 2D PAGE gels were<br />
superposed and comparative detailed analysis was performed. Accord<strong>in</strong>g to the analysis,<br />
37 prote<strong>in</strong> spots were differentially expressed. Among them, 5 prote<strong>in</strong>s were newly<br />
formed, 10 prote<strong>in</strong>s lost, 12 prote<strong>in</strong>s were up-regulated and 10 prote<strong>in</strong>s were down-<br />
regulated as compared to control group (untreated cells). These differentiated prote<strong>in</strong>s<br />
(<strong>in</strong>creased and/or decreased or appear and/or disappear) were recovered from gel to be<br />
<strong>applied</strong> for MALDI-TOF Mass Spectrometry for prote<strong>in</strong> identification and their<br />
function was evaluated. While sixteen prote<strong>in</strong>s were not identified because of some<br />
personal and <strong>in</strong>strumental errors, twenty one of them were determ<strong>in</strong>ed (it is considered<br />
that there were not seen two or more than peptide matches <strong>in</strong> the results). Accord<strong>in</strong>g to<br />
results, six prote<strong>in</strong>s are related with cell signal<strong>in</strong>g (Spot 7, 11, 14, 15, 23, 29), one<br />
prote<strong>in</strong> has a role <strong>in</strong> cell survival (Spot 34), one is cell cycle regulator (Spot 4), two<br />
prote<strong>in</strong>s are <strong>in</strong>volved <strong>in</strong> apoptosis (Spot 2, 27), one prote<strong>in</strong> is concern<strong>in</strong>g tumor<br />
suppressor (Spot 8),five prote<strong>in</strong>s take a key role <strong>in</strong> immunity and defense mechanism of<br />
the body (Spot 9, 12, 18, 25, 35), one prote<strong>in</strong> is <strong>in</strong>volved <strong>in</strong> prote<strong>in</strong> modification and<br />
prote<strong>in</strong> ubiquit<strong>in</strong>ation (Spot 22) and three of them has other functions (Spot5, 6, 10).<br />
For the further step studies, the same anti-cancer agent can be tried to treat other<br />
cancer types, with the help of differentially expressed prote<strong>in</strong>s. Additionally, the data<br />
obta<strong>in</strong>ed by this study can be used to improve new treatments for Multiple Myeloma by<br />
drug designers.<br />
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