A novel dysfunctional p53 mutation in the human neuroblastoma cell line TGW.

Mutations of p53 are rare in primary and advanced neuroblastomas. The p53 gene was studied in a TGW cell line established from a TNB1 xenograft, derived from metastasized neuroblastoma. The p53 protein level in TGW was elevated at baseline.

Multiple sites required for expression in 5'-flanking region of the hMLH1 gene.

Expression of the hMLH1 gene, one of the DNA mismatch repair genes, is frequently repressed in various cancers such as colorectal, ovarian, gastric, and endometrial origins with a microsatellite instable phenotype. In this study, we investigated details of the relationship between the transcriptional activity and the protein-binding sites in the 5'-flanking region of the hMLH1 gene. Luciferase reporter gene analysis with a series of deletion mutants revealed that a region containing -301 to -76 relative to a translation start site is essential for maximal expression.

Angiotensin-converting enzyme (ACE) I/D genotype and renal ACE gene expression.

Background: The angiotensin-converting enzyme (ACE) I/D genotype affects serum ACE levels and the onset and progression of renal disease, but little is known about the mechanism. We investigated a possible association between the ACE I/D genotype and renal ACE mRNA levels in healthy subjects.

Methods: Renal biopsy samples were obtained from 50 healthy kidney donors.

Various AGC repeat numbers in the coding region of the human transcription factor gene E2F-4.

The E2F family of transcription factors regulates the expression of genes required for DNA synthesis and cell cycle control. The AGC triplet repeat in the coding region of the E2F-4 gene, a member of the family, has been reported to be mutated in colorectal cancers with a microsatellite instability (MSI) phenotype. We found a wider range variation of the repeat number in DNAs from tumors, the corresponding normal mucosa, and healthy individuals.

A common polymorphism in exon 40 of the human mannose 6-phosphate/insulin-like growth factor II receptor gene.

Mammalian mannose 6-phosphate/insulin-like growth factor II receptor (M6P/IGF2R) binds insulin-like growth factor II and ligands containing an M6P recognition marker like the latent transforming growth factor-beta. Its genetic alteration has been associated with tumorigenesis of various tumours. We report here a novel polymorphism in exon 40 of the human M6P/IGF2R gene (6206A-->G, Asn2020Ser).

Molecular analysis and diagnosis in Japanese patients with Wilson's disease.

Background: Wilson's disease is characterized by the toxic accumulation of copper in the liver, brain, cornea and other organs. It is caused by both impaired excretion via the bile and impaired incorporation of copper into ceruloplasmin in the liver. The Wilson's disease gene (ATP7B) has been cloned as a putative copper-transporting P-type ATPase gene.

Competitive polymerase chain reaction for the determination of N-myc amplification in neuroblastoma: report of clinical cases.

If an unfavorable prognosis is suspected in neuroblastoma, decision on a treatment protocol should be based on the N-myc copy number (12). We already demonstrated that the newly developed competitive polymerase chain reaction (competitive PCR) is a promising method for the determination of the N-myc copy number (6), and have started to use this competitive PCR procedure in neuroblastoma patients, together with fine-needle biopsy in selected cases. Seven children were studied.

Quantitation of c-erbB-2 gene amplification in breast cancer tissue by competitive PCR.

Controversy exists regarding the relationship of the degree of c-erbB-2 amplification to other prognostic factors in breast cancer. To determine the degree of amplification of c-erbB-2 exactly, a sensitive and quantitative method is required. We have developed a competitive PCR method to quantitatively determine the amplification of the c-erbB-2 oncogene.

Extensive genetic heterogeneity in the neuroblastoma cell line NB(TU)1.

A neuroblastoma cell line displaying genetically unique features was established from a stage III case of a 20-month-old girl. Southern blotting by the probe pTNB6, which contains exon 1 of the N-myc gene, showed that the primary tumor had in total 4 aberrant bands beside the normal amplified band. The established cell line NB(TU)1 had an aberrant N-myc band (9.

Undectable expression of hMLH1 protein in sporadic colorectal cancer with replication error phenotype.

Purpose: Four DNA mismatch repair genes have been identified as being susceptible genes for hereditary nonpolyposis colorectal cancer. Deficiency of one of the mismatch repair genes causes the replication error phenotype in more than 80 percent of patients with hereditary nonpolyposis colorectal cancer and in 10 to 30 percent of patients with sporadic colorectal cancer. To determine which mismatch repair gene is lacking the function in patients with replication error-positive colorectal cancer, several approaches have been used at the nucleic acid and protein levels.

Competitive polymerase chain reaction for the quantification of N-myc gene copy number in neuroblastoma.

An absolute quantification method for the N-myc gene copy number of neuroblastoma specimens was established by applying the competitive polymerase chain reaction (cPCR). The competitor plasmid (pZH2) lacking an MluI site in the exon 2 was constructed to distinguish two product species amplified from genomic DNA and the competitor plasmid. By using this cPCR system, we could obtain qualitative results within 1 day, i.

Genetic clinical markers of human neuroblastoma with special reference to N-myc oncogene: amplified or not amplified?--An overview.

Neuroblastoma is the most common extracranial tumor in children, and cytogenetically, chromosome 1p deletions, extrachromosomal double minutes, and homogeneously staining regions (HSRs) are commonly observed in cell lines and in tumors in advanced stages. It is found that an HSR represents genomic amplification of N-myc, which plays a key role in determining the aggressiveness of neuroblastoma. However, stage IV neuroblastomas or cell lines which lack N-myc amplification are also progressive, and some of them show evidence of N-myc expression in terms of mRNA and/or N-Myc oncoprotein.

A novel RNA splicing mutation in Japanese patients with Wilson disease.

Deletion/insertion mutation of Wilson disease (WD) gene in 16 Japanese patients with Wilson disease was studied. A truncated size in a region of exon 4 to 6 was found by reverse transcription-polymerase chain reaction (RT-PCR) covering entire 21 exons except exon 1 for liver cDNA of one patient with a late onset neurologic type. Sequence analysis of the cDNA revealed that this truncation was occurred by skipping of exon 5, though any mutation in exon 5 of genomic DNA was failed to detect.

Coexpression of the myc gene family members in human neuroblastoma cell lines.

Members of the myc oncogene family such as c, N-, and L-myc are expressed in many malignant tumors. Expression of c-, N-, and L-myc oncogenes in 7 human neuroblastoma cell lines (GOTO, IMR-32, TGW, SCCH-26, TNB 9, NBL-S, and SK-N-SH), a human small cell lung carcinoma SBC-5 cell line, and a human monocytic leukemia THP-1-S cell line at mRNA and protein levels was studied to know the specificity of a newly developed antibody against homologous region at C-terminus of N-Myc, designated as anti pan-Myc antibody. By RT-PCR and immunoblot analysis, coexpression of three myc genes was detected in all neuroblastoma cell lines tested.

Angiotensin-converting enzyme polymorphism and development of diabetic nephropathy in non-insulin-dependent diabetes mellitus.

We determined the distribution frequency of angiotensin-converting enzyme insertion/deletion (I/D) polymorphism in 111 Japanese patients with non-insulin-dependent diabetes mellitus (NIDDM) of at least 10 years duration (80 patients with diabetic nephropathy and 31 patients without nephropathy) and 76 healthy Japanese controls. Patients with diabetic nephropathy showed an excess of the ID genotype compared with patients without nephropathy (p < 0.02) and less of the II genotype compared with healthy controls (p < 0.

Deficiency of holo-, but not apo-, ceruloplasmin in genetically copper-intoxicated LEC mutant rat.

Long-Evans Cinnamon(LEC) mutant rats exhibited less than 5% of normal levels of serum ceruloplasmin oxidase activity, but immunoblot analysis showed normal levels of immunologically detectable ceruloplasmin protein in sera from the mutant rats. Immunostaining of cryosections from the liver tissues with anti-ceruloplasmin antibody showed no significant difference between normal and LEC rats. Results from pulse labeling of ceruloplasmin for 3 hours with [35S]methionine in primary hepatocyte culture, followed by immunoprecipitation, SDS-PAGE and fluorography, showed only minor changes in ceruloplasmin protein synthesis and secretion.

Cloning of a Bacteroides gingivalis outer membrane protein gene in Escherichia coli.

Gene banks of chromosomal DNA from Bacteroides gingivalis 381 were constructed using the bacteriophage replacement vector lambda L47.1. A clone encoding an outer membrane protein from B.

The N-myc gene product in primary retinoblastomas.

The N-myc gene product in retinoblastomas was examined using the antisera against the N-myc gene product, which was produced as a fusion protein by Escherichia coli. The N-myc gene product was detected not only in the retinoblastoma cell line Y79 but also in primary retinoblastomas as a pair of bands of approximately 62 kilodaltons (KD) by immunoblotting. Immunohistochemical analysis showed positively stained cells with the antibody against the N-myc gene product in a few rosettes or fleuretts containing area of the tumor.

The primary structure of the leu1+ gene of Schizosaccharomyces pombe.

A DNA fragment which carries the leu1 gene encoding beta-isopropylmalate dehydrogenase in Schizosaccharomyces pombe has been isolated by complementation of an E. coli leuB mutation. This 1.