Detailed Information on Publication Record
2009
Combination of mRNA and protein microarray analysis in complex cell profiling
ŠIMARA, Pavel, Irena KOUTNÁ, Stanislav STEJSKAL, Petr KRONTORÁD, Zdeněk RUČKA et. al.Basic information
Original name
Combination of mRNA and protein microarray analysis in complex cell profiling
Name in Czech
Kombinace mRNA a proteinové microarray analýzy v komplexním profilování buněk
Authors
ŠIMARA, Pavel (203 Czech Republic, belonging to the institution), Irena KOUTNÁ (203 Czech Republic, guarantor, belonging to the institution), Stanislav STEJSKAL (203 Czech Republic, belonging to the institution), Petr KRONTORÁD (203 Czech Republic, belonging to the institution), Zdeněk RUČKA (203 Czech Republic, belonging to the institution), Martina PETERKOVÁ (203 Czech Republic, belonging to the institution) and Michal KOZUBEK (203 Czech Republic, belonging to the institution)
Edition
Neoplasma, Bratislava, Slovensko, AEPress, s.r.o. 2009, 0028-2685
Other information
Language
English
Type of outcome
Článek v odborném periodiku
Field of Study
10601 Cell biology
Country of publisher
Czech Republic
Confidentiality degree
není předmětem státního či obchodního tajemství
References:
Impact factor
Impact factor: 1.192
RIV identification code
RIV/00216224:14330/09:00034277
Organization unit
Faculty of Informatics
UT WoS
000263775100010
Keywords in English
microarray; cell profiling; protein expression; mRNA expression; HL-60
Tags
International impact, Reviewed
Změněno: 2/3/2018 09:49, Mgr. Pavel Šimara, Ph.D.
V originále
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.
In Czech
Tato studie kombinuje mRNA a proteinovou analýzu za využití cDNA microarrays a protilátkových microarrays. Integrací těchto dvou technologií získáváme komplexní pohled na buněčný molekulární profil. Studie byla provedena na leukemické buněčné linii HL-60, která diferencovala do granulocytárního stádia po přidání kyseliny retinové (ATRA). Tato buněčná linie tedy slouží jako in-vitro model hematopoezy. Pozorovali jsme změnu hladiny určitých proteinů, účastnících se zejména diferenciačních a apoptických signálních drah, často s onkogenetickým potenciálem. Hladina mRNA vždy neodpovídala hladině exprimovaného proteinu.
Links
| MSM0021622430, plan (intention) |
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| 2B06052, research and development project |
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