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@article{798695,
author = {Šimara, Pavel and Koutná, Irena and Stejskal, Stanislav and Krontorád, Petr and Ručka, Zdeněk and Peterková, Martina and Kozubek, Michal},
article_location = {Bratislava, Slovensko},
article_number = {56},
keywords = {microarray; cell profiling; protein expression; mRNA expression; HL-60},
language = {eng},
issn = {0028-2685},
journal = {Neoplasma},
title = {Combination of mRNA and protein microarray analysis in complex cell profiling},
url = {http://www.neoplasma.sk/},
volume = {2/2009},
year = {2009}
}
TY - JOUR
ID - 798695
AU - Šimara, Pavel - Koutná, Irena - Stejskal, Stanislav - Krontorád, Petr - Ručka, Zdeněk - Peterková, Martina - Kozubek, Michal
PY - 2009
TI - Combination of mRNA and protein microarray analysis in complex cell profiling
JF - Neoplasma
VL - 2/2009
IS - 56
SP - 141-149
EP - 141-149
PB - AEPress, s.r.o.
SN - 00282685
KW - microarray
KW - cell profiling
KW - protein expression
KW - mRNA expression
KW - HL-60
UR - http://www.neoplasma.sk/
N2 - This study combines mRNA and protein analysis using cDNA and antibody microarray techniques, respectively. These create a novel, integrated perspective into cellular molecular profiles. The aims of this study were to establish a reliable way of integrating these two approaches in order to obtain complex molecular profiles of the cell and to find suitable methods to normalize the data obtained using these approaches. Antibody microarray and cDNA microarray techniques were used to study expression alterations in HL-60 cells that were differentiated into granulocytes using all-trans retinoic acid (ATRA). We selected this model to evaluate this combined profiling technique because the expression levels of most of the mRNA and protein species in these cells are not altered; therefore it is easier to track and define those species that are changed. The proteins whose levels were altered included cmyc, c-jun, Pyk2, FAK, PKC, TRF1, NF-kappaB and certain caspase types. These proteins are involved in apoptosis and hematopoietic differentiation pathways, and some have also been reported to have oncogenic potential. We compared the results obtained using the two methods, verified them by immunoblotting analysis, and devised normalization approaches. This is one of the first demonstrations that a combination of antibody microarray and cDNA microarray techniques is required for complex molecular profiling of cells based on multiple parameters. This approach allows a more detailed molecular phenotype of the given sample to be obtained. The results obtained using a combination of the two profiling methods are consistent with those from previous studies that used more traditional methods.
ER -
ŠIMARA, Pavel, Irena KOUTNÁ, Stanislav STEJSKAL, Petr KRONTORÁD, Zdeněk RUČKA, Martina PETERKOVÁ a Michal KOZUBEK. Combination of mRNA and protein microarray analysis in complex cell profiling. \textit{Neoplasma}. Bratislava, Slovensko: AEPress, s.r.o., 2009, roč.~2/2009, č.~56, s.~141-149. ISSN~0028-2685.
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