Related Experiment Videos
Alteration in p53 modulates glial proteins in human glial tumour cells
1Division of Neurological Surgery, School of Medicine, University of California, San Diego, USA. hoisang@ucsd.edu
Abstract:
In transformed human glial cells, abnormalities of the p53 gene and altered expression of glial-specific properties (GSPs) have been observed. We therefore investigated whether (i) expression of the altered p53 protein is involved in the reduced expression of GSPs; and (ii) expression of the wild-type p53 (wt-p53) gene leads to induction of GSPs. We first determined that the p53 gene is mutated in human glioblastoma U-373MG cells. In these cells, and in human T-98G glioblastoma cells reported to possess a mutated p53 (m-p53) gene, nuclear m-p53 expression was intense while GSP expression was low in the same cell as revealed by double labelling immunocytochemistry. Conversely, glial fibrillary acidic protein (GFAP) and glutamate synthase (GS) were expressed in cells devoid of nuclear m-p53 immnunoreactivity. Therefore, a mutually exclusive relationship exists between the cytoplasmic GSPs and nuclear m-p53. Upon treatment with retinoic acid (RA) and dibutyryl cyclic AMP (dbcAMP), overall GSP staining were increased concomitant with suppression of nuclear m-p53. Their mutually exclusive expression pattern was maintained suggesting a functional relationship. This is supported by the observation of a similar mutually exclusive expression pattern for p53 and GSPs in pathologic specimens of human glioblastoma tissues. We then explored the role of the wt-p53 gene in the induction of GSPs using a wt-p53 tetracycline-regulated conditional expression system in human LN-Z308 glioblastoma cells. These cells normally express no p53 and no appreciable levels of GS or GFAP. Induced expression of wt-p53 lead to induction of GSP. These observations are consistent with the hypotheses that (i) nuclear m-p53 expression and cytoplasmic expression of GFAP and GS are inversely correlated, and (ii) expression of the wt-p53 gene leads to the expression of GSPs.
Insights
Mutated p53 protein expression inversely correlates with glial-specific properties (GSPs) in glioblastoma cells. Wild-type p53 gene expression, however, induces GSPs, suggesting a critical role in glial cell differentiation.
Area of Science:
- Neuroscience
- Molecular Biology
- Oncology
Background:
- Transformed human glial cells exhibit p53 gene abnormalities and altered glial-specific properties (GSPs).
- The relationship between p53 gene status and GSP expression in glioblastoma remains unclear.
Purpose of the Study:
- To investigate if mutated p53 (m-p53) expression reduces GSP levels.
- To determine if wild-type p53 (wt-p53) gene expression induces GSPs.
Main Methods:
- Utilized double labeling immunocytochemistry to assess p53 and GSP expression in glioblastoma cell lines (U-373MG, T-98G).
- Treated cells with retinoic acid (RA) and dibutyryl cyclic AMP (dbcAMP) to modulate GSP and p53 expression.
- Employed a tetracycline-regulated wt-p53 conditional expression system in LN-Z308 glioblastoma cells.
Main Results:
- A mutually exclusive expression pattern was observed between nuclear m-p53 and cytoplasmic GSPs (GFAP, GS) in glioblastoma cells.
- RA and dbcAMP treatment increased GSP expression while suppressing nuclear m-p53, maintaining their inverse correlation.
- Induced expression of wt-p53 in LN-Z308 cells led to the induction of GSPs.
Conclusions:
- Nuclear m-p53 expression is inversely correlated with cytoplasmic GFAP and GS expression in glioblastoma.
- Wild-type p53 gene expression promotes the expression of GSPs, suggesting a role in glial cell differentiation and tumor suppression.