Characterization and gene expression profiling in glioma cell lines with deletion of chromosome 19 before and after microcell‐mediated restoration of normal human chromosome 19

Abstract

Nearly 10% of human gliomas are oligodendrogliomas. Deletion of chromosome arm 19q, often in conjunction with deletion of 1p, has been observed in 65–80% of these tumors. This has suggested the presence of a tumor suppressor gene located on the 19q arm. Chromosome 19 deletion is also of interest due to the better prognosis of patients with deletion, including longer survival and better response to chemotherapy, compared with patients without deletion. Two glioma cell lines with deletion of 19q were used for chromosome 19 microcell-mediated transfer, to assess the effect of replacing the deleted segment. Complementation with chromosome 19 significantly reduced the growth rate of the hybrid cells compared with the parental cell lines. Affymetrix U133 Plus 2.0 Gene Chip analysis was performed to measure and compare the expression of the chromosome 19 genes in the chromosome 19 hybrid cell lines to the parental cell line. Probes were considered significantly different when a P value <0.01 was seen in all of the cell line comparisons. Of 345 probes within the commonly deleted 19q region, seven genes (APOE, RCN3, FLJ10781, SAE1, STRN4, CCDC8, and BCL2L12) were identified as potential candidate genes. RT-PCR analysis of primary tumor specimens showed that several genes had significant differences when stratified by tumor morphology or deletion status. This suggests that one or more of these candidates may play a role in glioma formation or progression.

Global extracellular vesicle proteomic signature defines U87-MG glioma cell hypoxic status with potential implications for non-invasive diagnostics

“…EVs from U87-MG, i.e. the most well-characterized human glioma cell-line [21, 22], grown under normoxic (EV NORM ) or hypoxic (EV HYP ) conditions were isolated by standard sequential ultracentrifugation [20]. The size distribution and morphology of EVs was analyzed by transmission electron microscopy (TEM), where EV NORM and EV HYP predominantly were found in the size range of 50–150 nm in diameter with no apparent difference in their morphology (Fig.…”

Quantitative expression analysis of the apoptosis‐related gene, BCL2L12, in head and neck squamous cell carcinoma

“…Therefore, it is tempting to speculate that BCL2L12 acts anti‐apoptotically in TSCC, one of the most aggressive types of HNSCC, resulting in the survival of cancerous squamous cells and the progression of the disease (55, 56). Similarly, recent oncogenomic studies uncovered the anti‐apoptotic role of the BCL2L12 oncoprotein in primary glioblastoma multiforme (GBM), where it is robustly expressed, at a significantly higher level than in oligodendrogliomas and oligoastrocytomas (57), and/or surrounding normal brain tissue (16). Furthermore, the BCL2L12 expression possesses significant discriminatory value, distinguishing efficiently patients with TSCC from non‐cancerous population, as demonstrated by the ROC analysis.…”

Clinical significance and novel mechanism of action of kallikrein 6 in glioblastoma

“…Additional, 10-mM frozen sections were then macrodissected on dry ice and the tumor tissue only placed in Trizol Reagent (Invitrogen) prior to RNA extraction. 29 Levels of KLK6 in RNA obtained from GBM were compared with those in 6 control nondiseased human total brain samples derived from 3 separate commercial sources (Ambion, Clontech, and Stratagene).…”