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Biomedical subjects

G Evan

Publications and source records attributed to G Evan.

52 records · Page 3Linked to original sources
PubMed1989

c-myc oncogene product expression and prognosis in operable breast cancer.

The 62 kDa protein product of the c-myc oncogene (p62 c-myc) is thought to be involved in the control of normal cellular proliferation and differentiation. We have measured oncoprotein levels using a flow cytometric assay in 141 operable breast cancers and have correlated levels with prognostic variables, patient survival and disease free intervals. High levels of p62 c-myc were associated with well differentiated tumours. There was no correlation with tumour DNA index, lymph node or oestrogen receptor status. C-myc oncoprotein levels were not predictive of patient survival or disease free interval. This relationship of oncoprotein levels with tumour histological grade is in keeping with the suggestion that the c-myc oncogene is important in the control of cellular differentiation. The other findings imply that measurement of c-myc oncoprotein levels does not yield useful prognostic information.

Breast Neoplasms↗
PubMed1989

Translocation of a store of maternal cytoplasmic c-myc protein into nuclei during early development.

The c-myc proto-oncogene is expressed as a maternal protein during oogenesis in Xenopus laevis, namely, in nondividing cells. A delayed translation of c-myc mRNA accumulated in early oocytes results in the accumulation of the protein during late oogenesis. The oocyte c-myc protein is unusually stable and is located in the cytoplasm, contrasting with its features in somatic cells. A mature oocyte contains a maternal c-myc protein stockpile of 4 x 10(5) to 6 x 10(5) times the level in a somatic growing cell. This level of c-myc protein is preserved only during the cleavage stage of the embryo. Fertilization triggers its rapid migration into the nuclei of the cleaving embryo and a change in the phosphorylation state of the protein. The c-myc protein content per nucleus decreases exponentially during the cleavage stage until a stoichiometric titration by the embryonic nuclei is reached during a 0.5-h period at the midblastula stage. Most of the maternal c-myc store is degraded by the gastrula stage. These observations implicate the participation of c-myc in the events linked to early embryonic development and the midblastula transition.

Animals↗
PubMed1989

[Role of c-myc protein in the early embryonic development of Xenopus].

Microinjections of antibodies directed against the protein encoded by the c-myc protooncogene strongly inhibit or arrest the early cell cleavage stage of Xenopus laevis embryos. Injections in one blastomere of a two cell stage embryo inhibit the segmentation of this blastomere. The cleavage of the uninjected blastomere behaves normally. Injections of control rabbit immunoglobulins do not alter the embryonic development.

Animals↗
PubMed1988

The human L-myc gene encodes multiple nuclear phosphoproteins from alternatively processed mRNAs.

The human proto-oncogene L-myc generates at least four different mRNAs by alternative RNA processing. We have identified two phosphorylated L-myc proteins with molecular masses of 60,000 and 66,000 daltons [p60L-myc(human) and p66L-myc(human)] in a small-cell carcinoma line expressing high levels of L-myc mRNA. These proteins have a short half-life and are localized to the nuclear matrix fraction, as previously reported for the c-myc and N-myc proteins. In vitro translation experiments demonstrated that both the p60 and p66 species are encoded by a 3.9-kilobase (kb) mRNA which retains intron 1, while only the p60 protein is translated from a 3.6-kb L-myc mRNA which has had intron 1 removed. While L-myc proteins [p32L-myc(human) and p37L-myc(human)] could be synthesized in vitro from 2.2-kb mRNA templates, no such proteins were detected by immunoprecipitation in vivo. These observations suggest that alternative RNA processing of the L-myc transcript could play a role in determining the steady-state levels of the p60L-myc and p66L-myc proteins.

Humans↗
PubMed1987

c-myc oncogene expression in colorectal cancer.

The pattern of c-myc gene organization and expression has been examined in resected colonic tumors and in the adjacent normal colon from 15 patients undergoing radical surgery. DNA hybridization showed no evidence of gene amplification or rearrangement. Transcripts of the c-myc messenger ribonucleic acid (mRNA) were elevated up to 32-fold in 12 of 15 tumors. The gene product, p62c-myc, was detected by both immunoblotting and immunohistology using a monoclonal antibody raised against a synthetic peptide immunogen. There was close correlation between c-myc mRNA copy number and p62c-myc abundance. Three well differentiated tumors contained high levels of transcript and protein, whereas four poorly differentiated tumors had the lowest levels. The assay of oncogene products may provide new biologically relevant tumor markers for determining prognosis and guiding treatment.

Aged↗
PubMed1987

Expression of c-myc in non-malignant and pre-malignant gastrointestinal disorders.

A monoclonal antibody 1-6E10 against the protein product p62c-myc of the c-myc oncogene was used to assess, by immunohistology, a variety of non-neoplastic and preneoplastic disorders of gastric and colonic mucosa. There were low levels of expression of the c-myc oncogenic product in normal gastric and colonic tissue. In gastric mucosa, increased expression was observed with inflammatory, metaplastic and dysplastic histological appearances. In the normal colon low levels of expression were observed, but there was increased expression in inflammatory disorders including Crohn's disease and ulcerative colitis. There was also increased expression in colonic dysplasia associated with ulcerative colitis. The oncogene product was localized in the cytoplasm, nuclei and Golgi apparatus. C-myc 1-6E10 may therefore be used as a marker to identify the cellular proliferative response in gastric and colonic mucosa that is associated with inflammation as well as potentially neoplastic hyperproliferative states.

Antibodies, Monoclonal↗
PubMed1987

A novel tumour marker related to the c-myc oncogene product.

We have studied the utility of the c-myc oncogene product as tumour marker using a set of monoclonal antibodies raised against synthetic peptides constructed from sequence data of the human c-myc oncoprotein. One antibody, Myc1-9E10, raised against the C-terminal 32 amino acids, has been shown to detect specifically the 62 kDa c-myc gene product in tumour cells. Immunoblotting of sera and urine with this antibody consistently revealed a single 40 kDa band (p40). Quantitative analysis using dilution dot immunoblotting demonstrated a considerable increase in the titre of p40 in the sera of 51 patients with a wide range of advanced solid tumours when compared with 17 healthy controls and 50 patients with non-malignant diseases. Serial measurement of the p40 titre in 12 patients with resected colorectal carcinoma shows a gradual return to normal with a half-life of 7 days. Our data suggests that p40 may be a useful new marker for monitoring tumour activity.

Biomarkers, Tumor↗
PubMed1987

Oncogenes and germ cell tumours.

The central problem in cancer therapy is the poor selectivity of current systemic agents against the common solid tumours. The demonstration that unique segments of DNA, constant in location and conserved in evolution are involved in growth control opens new avenues for basic and clinical research. The function of the products of these genes needs to be elucidated. Examples of growth control functions include homology to growth factors, surface receptors, protein kinases and cell cycle control proteins. From DNA sequence data peptides predicted to be exposed within intact molecules can be constructed and used to produce monoclonal antibodies to oncogene products. Such antibodies have now been successfully used to demonstrate the intracellular localization of gene products as well as the cell cycle regulatory role of the c-myc protein. By having a battery of antibodies against the different gene products their direct clinical application for diagnosis and prognosis has become a reality. Immunohistology and flow cytometry permit the geographical and quantitative analysis of function in normal and neoplastic tissues. Furthermore, by purification and biochemical analysis the molecular basis for their action can be elucidated. It is likely that by the end of the decade new drugs that inhibit oncoprotein function will be available for clinical trial.

Animals↗
PubMed1987

Expression of c-myc oncogene in coeliac disease.

A monoclonal antibody, produced by peptide immunisation was used to detect the distribution of p62c/myc by immunohistology in normal and coeliac small intestinal mucosa. The effect of gluten in four treated coeliac patients was investigated by taking serial jejunal biopsy specimens for six hours after a 10 g oral gluten challenge. There was a progressive increase in p62c/myc staining intensity in the villus enterocytes extending to the crypts, which accompanied the classical morphological changes occurring in the mucosa.

Antibodies, Monoclonal↗
PubMed1986

Raising antibodies by coupling peptides to PPD and immunizing BCG-sensitized animals.

The use of PPD (purified protein derivative of tuberculin) as a carrier has several significant advantages. It provides very powerful T cell help and it gives rise to virtually no antibody response against itself. This is particularly useful if it is intended to go on to make monoclonal antibodies, where the presence of a large amount of anti-carrier antibody is a nuisance! Furthermore, unlike most comparably powerful adjuvant systems, it can be used in man. PPD coupling has been used to raise antibodies to haptens and to raise T cell responses to tumour cells. It is here reported that small peptides coupled to PPD will give rise to good titres of anti-peptide antibody. For peptides that contain no cysteine, coupling has been achieved by attaching succinimidyl 4-(N-maleimidomethyl) cyclohexane-1-carboxylate (SMCC) to the alpha-amino group of the peptide and N-succinimidyl 3-(2-pyridyldithio) propionate (SPDP) to the PPD and allowing an uncleavable bond to form between them. Data on immunization with the leucotactic nonapeptide of the alpha chain of the complement component C3 and with some oncogene-related peptides have been obtained.

Animals↗
PubMed1986

Detection of the c-myc oncogene product in colonic polyps and carcinomas.

The c-myc oncogene has been implicated in the processes of normal cell proliferation and differentiation. Elevated levels of c-myc mRNA and its gene product (p62c-myc), have been detected in a variety of solid tumours and cultured cel lines. Its precise role in normal cell function and in neoplastic transformation and progression has yet to be elucidated. We have used a monoclonal antibody, raised by peptide immunisation, to determine the distribution by immunoperoxidase staining of the c-myc oncogene product in archival specimens of colonic polyps and carcinomas. Samples from 42 patients with colon carcinoma, 24 with benign polyps and 15 normal colon biopsies were examined. Normal colon revealed maximum staining in the mid-zone of the crypts, corresponding to the zone of differentiation and maturation. The staining was predominantly cytoplasmic. Adenomatous polyps revealed the most intense pattern of staining in areas of dysplastic change. Colonic tumours showed a wide range of staining. Well differentiated tumours contained more cytoplasmic p62c-myc than poorly differentiated tumours. These findings suggest that the c-myc oncogene product may play an important role in the evolution of colonic neoplasia.

Adenoma↗
PubMed1986

The distribution of the c-myc oncogene product in malignant lymphomas and various normal tissues as demonstrated by immunocytochemistry.

The expression of c-myc was studied in 51 malignant lymphomas and in a variety of normal tissues by immunocytochemistry using monoclonal antibodies raised to different synthetic peptides and reacting monospecifically with the c-myc product (p62c-myc). The c-myc product was detected in only a minority of malignant lymphomas principally those containing cells with immunoblastic characteristics, and was predominantly localised to the cytoplasm. In normal lymphoid tissues only plasma cells and histiocytes were found to have immunoreactivity. In non-lymphoid normal tissues, however, the c-myc product was distributed widely. Marked differences in its intracellular distribution were apparent in different tissues. These findings suggest that the relationship of p62c-myc expression to cell division may be more complex than previously suggested by in vitro studies, and raise the possibility that it may have other functions within the cell.

Antibodies, Monoclonal↗
PubMed1985

Metabolism of c-myc gene products: c-myc mRNA and protein expression in the cell cycle.

The presence and synthesis of c-myc protein and mRNA in the cell cycle has been studied. We find that c-myc mRNA is present, at equivalent levels, at all times in the cell cycle with the possible exception of mitosis. Furthermore, we demonstrate that this mRNA is transcribed in both G1 and G2 phases. An analysis of the c-myc protein in vivo shows that de novo synthesis occurs in G1 and G2 and the protein turns over with a half-life of approximately 20-30 min in both phases. Furthermore, the level of c-myc protein rapidly increases in cell populations when they re-initiate the cell cycle, thereafter decreasing as the culture reaches quiescence. The results therefore suggest that expression of c-myc can be rapidly modulated and that it is activated during the G0 to G1 transition, but is expressed thereafter in the cell cycle.

Animals↗
PubMed1985

Detection of the c-myc oncogene product in testicular cancer.

A set of monoclonal antibodies was constructed by immunising mice with peptide fragments of the c-myc oncogene product. One such antibody, Myc 1-6E10 was shown to bind to a 62,000 dalton protein identifiable with the c-myc product (p62c-myc). The antigen recognised was not destroyed by paraffin wax embedding. Myc 1-6E10 was used to characterise the distribution of p62c-myc in archival testicular tumour material. Normal testes expressed only small amounts of p62c-myc. Seminomas showed increased nuclear and cytoplasmic staining. Undifferentiated teratoma showed little activity, whereas p62c-myc was abundant in the nuclei of differentiating epithelial structures, yolk sacs and embryoid bodies. Only small amounts of p62c-myc were seen in the tumours of 5 patients who subsequently died from their disease.

Antibodies, Monoclonal↗
PubMedDate not supplied

Induction of c-fos-like protein in spinal cord neurons following sensory stimulation.

It has been suggested that the proto-oncogenes c-fos and c-myc participate in the control of genetic events which lead to the establishment of prolonged functional changes in neurons. Expression of c-fos and c-myc are among the earliest genetic events induced in cultured fibroblast and phaeochromocytoma cell lines by various stimuli including growth factors, peptides and the intracellular second messengers diacylglycerol, cAMP and Ca2+. We report here that physiological stimulation of rat primary sensory neurons causes the expression of c-fos-protein-like immunoreactivity in nuclei of postsynaptic neurons of the dorsal horn of the spinal cord. Activation of small-diameter cutaneous sensory afferents by noxious heat or chemical stimuli results in the rapid appearance of c-fos-protein-like immunoreactivity in the superficial layers of the dorsal horn. However, activation of low-threshold cutaneous afferents results in fewer labelled cells with a different laminar distribution. No c-fos induction was seen in the dorsal root ganglia, gracile nucleus or ventral horn. Thus, synaptic transmission may induce rapid changes in gene expression in certain postsynaptic neurons.

Animals↗