Detection of the PGP9.5 and Tyrosine Hydroxylase mRNAs for Minimal Residual Neuroblastoma Cells in Bone Marrow and Peripheral Blood.

Abstract

The "touchdown" polymerase chain reaction (PCR) technique has been applied to analyze expression of the neuron-specific protein, PGP9.5, and tyrosine hydroxylase (TH) genes for detection of minimal residual neuroblastoma cells in bone marrow and peripheral blood. PGP9.5 and TH gene products were not detected in any normal samples (n = 72) examined. However, in patients more than 1 year of age with stage III and IV neuroblastoma PGP9.5 mRNA was detected in six of seven bone marrow samples and in four of eight peripheral blood samples, and TH mRNA in four of seven and three of eight, respectively. The detection sensitivity was up to 10(-6) to 10(-7) micrograms of total cellular RNA for PGP9.5 and 10(-4) micrograms for TH. Among forty bone marrow specimens from nineteen patients with neuroblastoma both PGP9.5 and TH mRNAs were detected in six, and only PGP9.5 mRNA was detected in ten. Since detection of PGP9.5 and TH gene transcripts by the "touchdown" PCR was highly specific and sensitive, it might be most informative at present to carry out both PGP9.5 and TH mRNA assays for minimal residual neuroblastoma cells in blood and bone marrow.

Hypomethylation of the protein gene product 9.5 promoter region in gallbladder cancer and its relationship with clinicopathological features

“…as the cause of transformation. (19)(20)(21)(22)(23)(24)(25)(26)(27) Although PGP9.5 is known to be a dual function protein, that is, as a hydrolase to generate free ubiquitin and as a ligase to produce multi-ubiquitin chains, (17,41) both activities are involved in regulating ubiquitin levels. Moreover, the accumulation of ubiquitin has been documented in various types of primary cancers.…”

“…(27) Interestingly, a previous methylation analysis of the promoter region of PGP9.5 showed that a virtual absence of methylated CpG sites in lung cancer cell lines is consistent with relatively high PGP9.5 expression, and that dense methylation contributes to the lack of its expression in HeLa cells, (28) suggesting that PGP9.5 expression is regulated by promoter methylation. Here, we analyzed the methylation status of CpG islands in the promoter region of human PGP9.5 in resected primary GB cancers and searched for correlations between this and the clinicopathological features of this disease.…”

Activity-based ubiquitin-specific protease (USP) profiling of virus-infected and malignant human cells

“…Despite the lack of information on their substrate specificity, it is clear that some USPs exert distinct growth regulatory activities by acting as oncoproteins (12)(13)(14) or tumor suppressor proteins (15)(16)(17). Furthermore, overexpression of certain USPs correlates with progression toward a more malignant phenotype in neuroblastoma (18) and carcinomas of lung, kidney, breast, and prostate (19)(20)(21)(22)(23), and the expression of some USPs is induced on growth factor stimulation (24)(25)(26). In addition, a strong increase of USP activity was observed on overexpression of the c-myc oncogene in cells of Burkitt's lymphoma (BL)-like phenotype, which correlated with resistance to apoptosis induced by pharmacological inhibition of the proteasome (27).…”

The ubiquitin C-terminal hydrolase UCH-L1 promotes bacterial invasion by altering the dynamics of the actin cytoskeleton

“…In addition, attenuated strains of L. monocytogenes and S. enterica serovar Typhimurium have been used as delivery vectors for immunogenic antigens (HPV E7) or enzymes, such as cytosine deaminase, that convert pro-drug to toxic drugs in tumour cells (Cunningham and Nemunaitis, 2001;Nemunaitis et al, 2003;Brockstedt and Dubensky, 2008;Maciag et al, 2009). Our findings predict that the success of this tumour-targeting strategy will be at least in part dependent on the expression of UCH-L1, suggesting that it may be particularly indicated for tumour types, such as neuroblastoma (Yanagisawa et al, 1998), colon carcinoma (Yamazaki et al, 2002), prostate and breast carcinomas (Schumacher et al, 1994;Aumuller et al, 1999) where UCH-L1 is expressed at very high levels.…”

“…The ubiquitin C-terminal hydrolase UCH-L1 is physiologically expressed at high levels in testis, ovary and neuronal cells, and is strongly upregulated in metastatic tumours and tumour-derived cell lines, including neuroblastoma (Yanagisawa et al, 1998), colon carcinoma (Yamazaki et al, 2002), prostate and breast carcinomas (Schumacher et al, 1994;Aumuller et al, 1999), and Burkitt's lymphoma (Rolen et al, 2008). This DUB was shown to be involved in several processes associated with the malignant phenotype, such as rapid proliferation and the capacity to form colonies in soft agar (Rolen et al, 2008;Kim et al, 2009).…”