ANALYSIS OF CTCF, ITS ANALOGUE (BORIS) AND YB1 PROTEIN IN GLIOMA AND OSTEOSARCOMA
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Date
2012-08
Authors
SITI ZAWANI, MOHD RAMLI
Journal Title
Journal ISSN
Volume Title
Publisher
Pusat Pengajian Sains Perubatan Universiti Sains Malaysia
Abstract
Human malignant glioma (DBTRG-05MG) and osteosarcoma (U2OS) cell lines derived from clinical samples are established in the laboratory and characterized. The cell lines underwent serially subculture in standard culture media with some alteration. Both cell lines showed heterogeneous morphology under phase- contrast observation. Proteomics study were conducted to analyze the present of specific proteins. The aim was to
establish human glioma and osteosarcoma primary cultures from clinical tumor samples and characterized the biochemical features of the cultures. The establishment of the cell lines will be used to support the future gene regulation study of gliomas and osteosarcoma malignancy and progression. CTCF, BORIS and YB-1 have been implicated in tumorgenesis. In this study, the expression of CTCF, BORIS and YB-1
protein and mRNA in glioma (DBTRG-05MG) and osteosarcoma (U2OS) cell lines were analyzed. In vivo physical interaction between CTCF and YB1 in both glioma and osteosarcoma cell lines were studied. The expressions of CTCF, BORIS and YB-1 in osteosarcoma (U2OS) and glioma (DBTRG-O5MG) in comparison with their respective normal osteoblast (hFOB 1.19) and glial (SVG-p12) cell lines were examined. Nucleotides mutation for CTCF (exon 5) and YB1 (exon 7) were also examined. The level of CTCF mRNA was significantly higher in osteosarcoma compared to osteoblast cell line, while the level of YB-1 mRNA was similar in these cell lines. The expression of YB-1 mRNA was significantly elevated in glioma compared to glial cell line, whereas expression of CTCF mRNA was similar. The BORIS mRNA was detected only in osteosarcoma cell line. Nucleotides analysis showed undetected mutations at exon 4 and 5 (CTCF) and exon 7 (YB1) for both cancerous cell lines. Protein expression of CTCF revealed immunopositive for all cancerous and normal cell lines. YB1 protein expressions in glial, glioma and osteosarcoma cells was found to be immunopositive and anomalously migrated. Positive in vivo physical interaction of CTCF~YB1 protein in
osteosarcoma and glioma cell lines were reported. Different patterns of expression of CTCF, BORIS and YB1 mRNAs indicates differential involvement of these transcriptional factors in tumorgenesis of osteosarcoma and glioma. Positive in vivo physical interaction of CTCF~YB1 in the osteosarcoma and glioma cell lines supported previous reports on both factors in tumor development. Based on the results, the expression patterns of CTCF/ BORIS and YB1 and protein-protein interactions of CTCF~YB1 require further fundamental research on its biochemical and functional characterization. It could be used as potential future prognostic marker in glioma and osteosarcoma. In conclusion, the glioma and osteosarcoma cell lines establishment
method was optimized with basic characteristics and the CTCF and YB-1 protein and gene showed potential roles in glioma and osteosarcoma development.
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Keywords
molecular medicine