ŠIMARA, Pavel, Irena KOUTNÁ, Stanislav STEJSKAL, Petr KRONTORÁD, Zdeněk RUČKA, Martina PETERKOVÁ and Michal KOZUBEK. Combination of mRNA and protein microarray analysis in complex cell profiling. Neoplasma. Bratislava, Slovensko: AEPress, s.r.o., 2/2009, No 56, p. 141-149. ISSN 0028-2685. 2009.
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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
Original language English
Type of outcome Article in a journal
Field of Study 10601 Cell biology
Country of publisher Czech Republic
Confidentiality degree is not subject to a state or trade secret
WWW Odborný vědecký článek
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 cbia-web, cell profiling, HL-60, microarray, mRNA expression, protein expression
Tags International impact, Reviewed
Changed by Changed by: Mgr. Pavel Šimara, Ph.D., učo 67594. Changed: 2/3/2018 09:49.
Abstract
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.
Abstract (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)Name: Funkční a molekulární charakteristiky nádorových a normálních kmenových buněk - identifikace cílů pro nová terapeutika a terapeutické strategie
Investor: Ministry of Education, Youth and Sports of the CR, Functional and molecular characteristics of cancer and normal stem cells - identification of targets for novel therapeutics and therapeutic strategies
2B06052, research and development projectName: Vytipování markerů, screening a časná diagnostika nádorových onemocnění pomocí vysoce automatizovaného zpracování multidimenzionálních biomedicínských obrazů (Acronym: Biomarker)
Investor: Ministry of Education, Youth and Sports of the CR, Determination of markers, screening and early diagnostics of cancer diseases using highly automated processing of multidimensional biomedical images
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