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Iran J Basic Med Sci. 2011 Sep;14(5):428-35.

Gene Expression Quantification of Toll like Receptors 2, 4 and Co-molecules in Human Glioblastoma Cell Line (U87-MG): Toward a New In vitro Model of Inflammation.

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1
Department of Biochemistry, Faculty of Science, Payam Noor University of Mashhad, Iran ; Department of Stem cells and Developmental Biology, Royan Institute for Stem cell Biology and Technology, ACECR, Tehran, Iran.

Abstract

OBJECTIVES:

Pattern recognition receptors (PRRs) are the main part in the innate immune response. Human glioblastoma cell line (U87-MG) is an established adherent cell line model of this common cancer; due to genetic variations between individuals it is likely more suitable for investigating molecular aspects of innate immunity. Therefore, we undertook a novel characterization of the immune phenotype of U87-MG toward establishing a base for future researches.

MATERIALS AND METHODS:

In this study, U87-MG cells where cultured in a normal condition, to investigate levels of toll-like receptor 2 (TLR2), TLR4, myeloid differentiation factor-88 (MyD88) and CD14 transcripts expression in these cells. Both RT-PCR and qPCR were applied to detect and quantify the expression levels of these genes in U87-MG cells and compare them to their levels in the peripheral blood mononuclear cells (PBMC) of healthy individuals, as a common reference.

RESULTS:

Expression level of TLR2 and TLR4 are not significantly different in U87-MG cells in comparison to PBMC. Also, expression levels of MyD88 and CD14 in U87-MG cells are significantly lower than their levels in PBMC. Furthermore, expression levels of MyD88 and CD14 in both PBMC and U87-MG are significantly lower than TLR2 and TLR4 transcripts.

CONCLUSION:

The data reveal expression of TLR4, CD14, MyD88 and TLR2 genes in U87-MG cell line, for the first time. Expression detection of these genes in human glioblastoma cell line might have a potential for diagnosis of inflammatory mechanisms in immune mediated disorders of in vitro models of human brain inflammatory disease.

KEYWORDS:

Human glioblastoma cell line (U87-MG); Innate immunity; Quantitative RT-PCR Toll-like receptors

PMID:
23492674
PMCID:
PMC3586846
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