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Isolation and Flow Cytometric Analysis of Glioma-infiltrating Peripheral Blood Mononuclear Cells
Published on: November 28, 2015
Down-regulation of CD14 transcripts in human glioblastoma cell line U87 MG
Alireza Haghparast1, Maryam Heidari Kharaji, Amir Mohammad Malvandi
1Department of Pathobiology, Ferdowsi University of Mashhad, Mashhad, Iran. alireza.haghparast@gmail.com
Background:
Pattern recognition receptors (PRRs) are the main sensors of pathogen and danger signals in innate immunity of which Toll Like Receptors (TLRs) are the most studied ones. The contribution of PRRs in cerebral inflammation induced by microbial infection, tissue damage and cancer has not extensively been addressed so far. Glioma is the most common tumor of the central nervous system and glioblastomas are the most common and most malignant primary brain tumors.
Objective:
The objectives of the present study were to investigate the expression of several PRRs including TLR2, TLR4, MyD88 and CD14 transcripts in human glioblastoma cell line U87 MG and compare their expression level with peripheral blood mononuclear cells (PBMC) obtained from healthy individuals.
Methods:
Touchdown PCR (TD-PCR) and Real-time quantitative PCR (qPCR) were applied to detect and quantify the expression level of TLR2, TLR4, MyD88 and CD14 transcript in U87 MG cell line and (PBMC) of healthy individuals.
Results:
According to our results, human glioblastoma cell line U87 MG expresses TLR2, TLR4, MyD88 and CD14 transcripts in TD-PCR. Moreover, the quantification of the expression of these genes revealed a highly significant down-regulation of CD14 and a slight up-regulation of TLR2 transcripts as compared to PBMC of healthy individuals.
Conclusion:
The lower expression level of CD14 in human glioblastoma cell line, might have a potential implication for CD14 mediated cerebral pathology.