Tumor-specific alterations at the p53 gene locus were analyzed in 40 human brain tumor samples. Gliomas were more prevalent in young males and meningiomas in old females. Structural changes at the intron 1 region of the p53 gene were analyzed in these tumors by Southern blotting. Among the 40 tumors, 33 were informative and 21 of these (63.6%) informative cases showed loss of heterozygosity (LOH). This is the first report showing LOH at the intron 1 region of p53 gene in human brain tumors. The level of p53 mRNA, p53 protein and Ser 392 phosphorylated p53 protein were also analyzed in all tumor samples. Normal sized p53 mRNA and protein were present in all the tumor samples; however, their levels were 1.5- to 4-fold higher compared to the control suggesting deregulated p53 pathway in these tumors. No correlation was found between LOH status and the levels of p53 mRNA and protein. In all high-grade glioblastomas majority of the p53 protein existed as Ser 392 phosphorylated form as compared to low-grade gliomas. In addition, the percentage of Ser 392 phosphorylated form of p53 protein was lower in meningiomas and other brain tumor types irrespective of tumor grade. These results suggest involvement of Ser 392 phosphorylated form of p53 protein during the later stages of glioma development. These results also indicate that deregulation of p53 gene could occur at various steps in p53 pathway and suggest an overall deregulation of p53 gene in most brain tumor types.

A total of 40 human brain tumor samples were analyzed for tumor-specific alterations at the RB1 gene locus. Gliomas were more prevalent in younger males and meningiomas in older females. Southern blot analysis revealed loss of heterozygosity (LOH) at the intron 1 locus of RB1 gene in 19.4% of informative cases and this is the first report showing LOH at this locus in human brain tumors. Levels of RB1 mRNA and protein, pRb, and the percentage of hyperphosphorylated form of pRb were also analyzed in these tumors. Normal human fibroblast cell line WI38 was used as control in northern and western analysis. Normal sized RB1 mRNA and protein were present in all the tumor samples. Majority of the gliomas had 2.0-fold or higher levels of RB1 mRNA and most meningiomas had less than 2.0-fold of RB1 mRNA compared to control WI38 cells. The total pRb levels were 2.0-fold or higher in all the tumor samples compared to control. More than 50% of pRb existed in hyperphosphorylated form in all gliomas except two. However, six out of 13 meningiomas had less than 50% of total pRb in the hyperphosphorylated form. These results indicate that the increased percentage of hyperphosphorylated form of pRb in gliomas could provide growth advantage to these tumors. Presence of LOH at the RB1 gene locus and the increased levels of RB1 RNA and protein and increased percentage of hyperphosphorylated form of pRb are indicative of an overall deregulation of pRb pathway in human brain tumors.