J 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:

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
Změněno: 2/3/2018 09:49, Mgr. Pavel Šimara, Ph.D.

Abstract

ORIG CZ

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)
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 project
Name: 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
Displayed: 8/11/2024 14:44