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T J McDonnell

Publications and source records attributed to T J McDonnell.

At least 19 recordsLinked to original sources
PubMed1995

Regulation of the proto-oncogenes bcl-2 and c-myc by the Wilms' tumor suppressor gene WT1.

The Wilms' tumor gene WT1 functions as a tumor suppressor gene, repressing transcription of several growth factors and growth factor receptors. The bcl-2 and c-myc proto-oncogenes are essential for regulation of apoptosis and cell proliferation with roles in development and oncogenesis. We found that WT1 can repress transcription of both the bcl-2 and c-myc promoters. This suggests that WT1 regulates bcl-2 and c-myc during renal development, and the loss of functional WT1 results in deregulation of bcl-2 and c-myc, contributing to tumor formation.

Amino Acid Sequence↗
PubMed1995

Cell death regulation during multistep lymphomagenesis.

The three most common genetic abnormalities occurring in malignant lymphomas involve alterations resulting in the deregulated expression of the c-myc and bcl-2 oncogenes and the inactivation of the p53 tumor suppressor gene. Relevant strains of genetically engineered mice, including bcl-2-Ig and E mu-myc transgenic mice and p53 knockout mice, have been used to prospectively examine the regulation of apoptotic cell death by these genes, individually and in combination, and their contribution to in vivo lymphomagenesis. The potential importance of the therapeutic induction of apoptosis is discussed.

Animals↗
PubMed1995

Apoptosis induction mediated by wild-type p53 adenoviral gene transfer in squamous cell carcinoma of the head and neck.

Cancer gene therapy strategies for inducing apoptosis in solid tumors may allow contemporary medicine to reassess its management of these cancers. We demonstrated previously that overexpression of the wild-type p53 gene in squamous cell carcinoma of the head and neck cell lines via adenovirus-mediated gene transfer suppressed growth both in vitro and in vivo. Here, we characterize the mechanism of the growth suppression by the exogenous p53 gene as a consequence of programmed cell death (apoptosis). One of the cell lines used in this study, Tu-138, harbors a mutated p53 gene, whereas the other cell line, MDA 686LN, possesses a wild-type p53 gene. DNA fragmentation was detected by electrophoresis in both cell lines after infection with the wild-type p53 adenovirus, Ad5CMV-p53. With the use of the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labeling method, 4.4% of the remaining viable Tu-138 cell population was identified as apoptotic as early as 15 h after inoculation with Ad5CMV-p53. The percentage of apoptotic cells increased to 31% at 22 h. In contrast, only 10% of the viable MDA 686LN cells (wt-p53) had undergone apoptosis 30 h after Ad5CMV-p53 infection, although the percentage of apoptotic cells rapidly increased to 60% at 48 h after infection. For in vivo analysis of apoptosis, nude mice in which squamous cell carcinoma of the head and neck cell lines had been implanted s.c. had exogenous wt-p53 transiently introduced to the tumor cells via Ad5CMV-p53 2 days later. In situ end labeling clearly illustrated apoptosis in the tumor cells. These results suggest that wt-p53 plays an important role in the induction of apoptosis in human head and neck cancer cell lines and that selective induction of apoptosis in cancer cells can be further explored as a strategy for cancer gene therapy.

Adenoviridae↗
PubMed1995

Combination adriamycin and suramin induces apoptosis in bcl-2 expressing prostate carcinoma cells.

Bcl-2 expression has been associated with progression of prostate cancer from androgen-dependence to androgen-independence and may contribute to the relative drug-resistant phenotype typically observed in androgen-independent prostate cancer. Dunning-G rat prostate cancer cells transfected with a bcl-2 expression vector demonstrated resistance to apoptosis induced by adriamycin and, to a lesser extent, suramin. Use of adriamycin and suramin in combination, however, circumvents this bcl-2 associated drug resistance. Our findings indicate that combination drug actions may induce apoptosis in resistant malignant cell types with defective apoptotic pathways.

Animals↗
PubMed1995

Evidence that c-myc mediated apoptosis does not require wild-type p53 during lymphomagenesis.

Deregulation of c-myc, frequently implicated in oncogenesis, is associated with increased cell proliferation and also cell death. Similarly, the p53 tumor suppressor gene commonly mutated in human tumors, is known to induce apoptosis or cell cycle arrest in its wild-type conformation. Genetically altered mice simultaneously overexpressing c-myc and possessing a disrupted p53 gene were used to investigate whether c-myc mediated apoptosis requires wild-type p53. The accelerated development of malignant lymphomas in these mice was found to be a consequence of enhanced proliferation and not reduced apoptosis resulting from the synergistic effect of c-myc overexpression and p53 inactivation.

Alleles↗
PubMed1995

Flow cytometric analysis of apoptosis and bcl-2 in human solid neoplasms.

Apoptosis (programmed cell death) and the genes regulating this process (e.g., bcl-2), have recently become a focus of interest in the study of cancer development and progression. We adapted flow cytometric techniques for measuring apoptosis and bcl-2 protein in solid tissues and concomitantly determined both parameters in 65 malignant solid neoplasms. Four different fixation methods were evaluated for bcl-2 analysis using a lymphoma cell line possessing a t(14;18), and the Daudi cell line as positive and negative controls, respectively; optimal fixation was achieved using 70% ethanol. Apoptosis was determined using the terminal deoxynucleotidyl transferase (TdT) end-labeling method of (Gorczyca et al.: Cancer Res 53:1945-1951, 1993). Treated (5Gy radiation, 4 h) and untreated portions of a murine cell line were used as positive and negative controls for apoptosis induction. For tumor specimens, bcl-2 positivity ranged from 0 to 89.5% (15.8 +/- 22.9), and apoptosis (TdT end-labeling) ranged from 0.4 to 84.3% (15.9 +/- 17.0). Generally, tumors with high bcl-2 expression (> or = 20.0%) showed significantly lower numbers of apoptotic cells than those with low (< 20.0%) bcl-2 (P = 0.05). A subset of tumors, however, exhibited low values for both parameters. We also observed that the proliferative fractions of tumors with high apoptosis (> or = 15.0%) were significantly different from those with low (< 15.0%) apoptosis (P = 0.005); higher proliferative rates were associated with high apoptosis. We conclude that optimal bcl-2 analysis is achieved using ethanol fixation and that flow cytometry provides a rapid and reliable technique for the measurement of these parameters. Concurrent analysis of these markers provides detailed information on the biological characteristics of tumor subpopulations and may assist in categorizing tumors for different management strategies.

Apoptosis↗
PubMed1995

Induction of apoptotic cell death in non-melanoma skin cancer by interferon-alpha.

Interferon-alpha (IFN-alpha) is a cytokine that is effective in the treatment of a variety of cancers, including non-melanoma skin cancers. The biologic responses of cells to IFN-alpha are pleiotropic and include growth suppression and immunomodulation. The potential direct effects of IFN-alpha on tumor cell populations are incompletely characterized. Our findings indicate that IFN-alpha can directly induce apoptosis (programmed cell death) in sensitive squamous cell skin cancer cell lines. Cell lines resistant to the cytotoxic effects of IFN-alpha showed no evidence of apoptosis induction. Transfection of IFN-alpha-sensitive cell lines with a bcl-2 expression vector conferred partial resistance to cell death induction by IFN-alpha. Our results indicate that the clinical efficacy of IFN-alpha may, in part, be related to the ability of this cytokine to induce apoptosis.

Apoptosis↗
PubMed1995

The functional basis of c-myc and bcl-2 complementation during multistep lymphomagenesis in vivo.

Oncogenes are known to be deregulated by chromosomal translocations occurring at high frequency in specific malignancies. Among the most well characterized of these are c-myc, associated with the t(8;14) in Burkitt's lymphomas, and bcl-2, associated with the t(14;18) in follicular lymphomas. In addition to their role in regulating rates of proliferation, it is known that oncogenes and tumor suppressor genes can also regulate rates of apoptotic cell death. The contribution of c-myc and bcl-2 to the regulation of cell death during lymphomagenesis in vivo is assessed using bcl-2-Ig and emu-myc trangenic mice and bcl-2/myc hybrid transgenic mice. Translocations between the endogenous c-myc gene and immunoglobulin loci, e.g., t(12;15), are common in lymphomas arising in the bcl-2-Ig mice. Furthermore, bcl-2/c-myc double transgenic mice exhibit accelerated lymphomagenesis, indicating cooperation between these two oncogenes. Genetic complementation of c-myc and bcl-2 during lymphomagenesis resulted from the suppression of c-myc-associated apoptosis. Other genes are likely involved in regulating cell death during multistep lymphomagenesis.

Animals↗
PubMed1995

Implications of apoptotic cell death regulation in cancer therapy.

The process of programmed cell death, or apoptosis, is now recognized as an important component of multistep carcinogenesis. Traditionally, interventions used in the treatment of cancer patients were predicated on enhanced rates of proliferation in the tumor cells. It is now appreciated that tumor growth may, in many circumstances, be more a function of reduced rates of cell death rather than enhanced proliferation per se. Our understanding of the molecular regulation of apoptotic cell death is leading to the design and implementation of treatment strategies based on cell death induction.

Animals↗
PubMed1995

The expression and localization of bcl-2 protein in normal skin and in non-melanoma skin cancers.

Non-melanoma skin cancers are the most commonly diagnosed malignancies and are typically indolent in their clinical behavior. Although predisposing factors leading to the development of these cancers, such as ultraviolet irradiation, are well described, the molecular events involved in their pathogenesis are incompletely understood. The localization of bcl-2 expression within the skin was determined using immunohistochemical methodologies and an anti-bcl-2 monoclonal antibody. The cytoarchitectural distribution of bcl-2 protein in normal skin included basal keratinocytes, the dermal papillae of the hair follicle, the keratinized Huxley's and Henle's layers, and the keratinized outer root sheath cells of the isthmus and infundibulum of the hair follicle. In addition, intense immunoreactivity was noted in the secretory coil of eccrine sweat glands. The distribution of bcl-2 protein within normal skin did not correlate with the known histologic localization of stem cell compartments. Basal cell carcinomas expressed high levels of bcl-2 protein. In contrast, squamous cell carcinomas typically exhibited no immunohistochemically detectable bcl-2 protein. The findings suggest a potential contribution of bcl-2 gene deregulation to the pathogenesis of some types of non-melanoma skin cancer.

Basal Cell Carcinoma↗
PubMed1994

Apoptosis and expression of the bcl-2 proto-oncogene in the fetal and adult human kidney: evidence for the contribution of bcl-2 expression to renal carcinogenesis.

bcl-2 was identified as a transcript associated with the t(14;18) and imparts resistance to apoptotic cell death. bcl-2 is normally expressed in many tissues and exhibits remarkable structural and functional conservation. Using immunohistochemical and in situ DNA labeling techniques we examined the localization of bcl-2 in the developing human kidney. bcl-2 expression was rapidly upregulated in the induced metanephrogenic mesenchymal cells that differentiate into renal vesicles and nephrons. bcl-2 expression was undetectable in uninduced mesenchyme and in the renal ampullae and associated collecting system. The distribution of apoptotic cells within the developing kidney was inversely correlated with expression of bcl-2. The localization of bcl-2 protein in the adult human kidney also was examined. bcl-2 was expressed at high levels in all renal neoplasms examined providing a potential basis for the deregulation of apoptosis in the development and progression of these tumors.

Adult↗
PubMed1994

Pseudomonas aeruginosa exotoxin A induces pulmonary endothelial cytotoxicity: protection by dibutyryl-cAMP.

In pseudomonal septicaemia, serum levels of antibody to exotoxin A have been demonstrated to be an important independent predictor of survival. Previously, we have demonstrated that exotoxin A directly injures pulmonary endothelial cells, and that dibutyryl-cyclic adenosine monophosphate (Db-cAMP) can attenuate this injury. The object of this study was to examine the mechanisms of this pulmonary endothelial cell injury and the mechanism of Db-cAMP protection. The effects of differing duration of exposure to exotoxin A and a reduction in temperature on endothelial cell injury were examined. In addition, the effect of post-treatment with Db-cAMP on exotoxin A-induced endothelial cell injury was studied. A brief, 5 min, exposure to exotoxin resulted in maximum injury comparable to that produced by 18 h exposure. This injury did not occur at low temperatures, which would inhibit receptor-mediated endocytosis. Db-cAMP protected endothelial cells, even when added up to one hour after exotoxin exposure. These results suggest that, in this model, exotoxin A-induced injury of endothelial cells is receptor-mediated. Furthermore, this injury may be attenuated even after exotoxin A internalization has taken place.

ADP Ribose Transferases↗
PubMed1992

Expression of the protooncogene bcl-2 in the prostate and its association with emergence of androgen-independent prostate cancer.

The significance of apoptosis in relation to the development and progression of prostate cancer remains largely undefined. bcl-2 is an oncogene that functions by overriding apoptosis. bcl-2 expression was localized to the basal epithelial cells in the normal human prostate with the use of immunohistochemistry. Androgen-dependent and androgen-independent prostate carcinomas were evaluated immunohistochemically for bcl-2 expression. bcl-2 was undetectable in 13 of 19 cases of androgen-dependent cancers. In contrast, androgen-independent cancers displayed diffuse, high levels of bcl-2 staining (P < 0.01). In rats, steady-state levels of bcl-2 mRNA, assessed by S1 assays, reached maximum levels 10 days following castration. Addition of exogenous testosterone with, or without, flutamide demonstrated that the increased bcl-2 mRNA resulted from androgen ablation. Our findings indicate that bcl-2 expression is augmented following androgen ablation and is correlated with the progression of prostate cancer from androgen dependence to androgen independence.

Androgens↗
PubMed1991

Bcl-2 maintains B cell memory.

The number of lymphocytes in an animal is remarkably constant despite antigen-driven proliferation and a high rate of B-cell lymphopoiesis. This reflects the relatively brief lifespan of many newly generated B cells and argues for a well-regulated death mechanism. Even so, a secondary immune response can be generated years after a primary exposure to antigen. Antigen that might restimulate B cells persists for extended periods on follicular dendritic cells in the light zone of germinal centres. Antigen-binding B cells have also been found months after the end of obvious cell division. The precise signal that enables certain B cells to emerge as long-term surviving memory cells is unknown. Bcl-2, an inner mitochondrial membrane protein, blocks programmed cell death in B cells. We report here that this proto-oncogene maintains immune responsiveness. Transgenic mice overproducing Bcl-2 have a long-term persistence of immunoglobulin-secreting cells and an extended lifetime for memory B cells.

Animals↗
PubMed1991

Progression from lymphoid hyperplasia to high-grade malignant lymphoma in mice transgenic for the t(14; 18).

Follicular lymphoma, the most common human lymphoma, characteristically has a t(14; 18) interchromosomal translocation. It is typically an indolent disease comprised of small resting B cells, but frequently develops into a high-grade lymphoma. The t(14; 18) translocates the Bcl-2 gene, generating a deregulated Bcl-2-immunoglobulin fusion gene. Bcl-2 is a novel inner mitochondrial membrane protein that extends the survival of certain cells by blocking programmed cell death. To determine the oncogenic potential of the t(14; 18) translocation, we produced transgenic mice bearing a Bcl-2-immunoglobulin minigene that structurally mimicked the t(14; 18). An indolent follicular hyperplasia in these transgenic mice progressed to a malignant diffuse large-cell lymphoma. The long latency, progression from polyclonal to monoclonal disease, and histological conversion, are all suggestive of secondary changes. Half of the immunoblastic high-grade lymphomas had a rearranged c-myc gene. Our transgenic mice provide an animal model for tumour progression in t(14; 18) lymphoma and show that prolonged B-cell life increases tumour incidence.

Animals↗
PubMed1990

Deregulated Bcl-2-immunoglobulin transgene expands a resting but responsive immunoglobulin M and D-expressing B-cell population.

We characterized the basis for the follicular lymphoproliferation in transgenic mice bearing a Bcl-2-immunoglobulin (Bcl-2-Ig) minigene representing the t(14;18) of human follicular lymphoma. Discriminatory S1 nuclease protection assays revealed that the Bcl-2-Ig transgene was overexpressed relative to endogenous mouse Bcl-2 in spleen and thymus. Western (immunoblot) analysis demonstrated the overproduction of the human 25-kilodalton Bcl-2 protein, which arose from the transgene, in spleen, thymus, and the expanded B-cell subset. Despite the generalized lymphoid pattern of deregulation, two-color flow cytometry and density gradient centrifugation indicated that the expanded lymphocytes were predominantly small, resting B cells coexpressing B220, immunoglobulin M (IgM), IgD, Ia, and kappa. Cell cycle analysis confirmed that about 97% of these expanded B cells reside in G0/G1. An extensive characterization of transgenic lines revealed a fourfold excess of IgM-IgD-expressing B cells in spleen and dramatically increased numbers in bone marrow. While resting, these cells proliferated in response to lipopolysaccharide and anti-IgM and demonstrated normal B-cell colony formation in soft agar. Moreover, these B cells, which demonstrated an extended survival in vitro even in the absence of stroma, were also resting in G0, yet were capable of proliferative responses. These findings provide consistent evidence that the accumulation of B cells after Bcl-2 overproduction is secondary to prolonged cell survival and not increased cell cycling. This suggests a unique role for Bcl-2 as a proto-oncogene that enhances cell survival independent of promoting cell division.

Animals↗
PubMed1990

Role of peptidoleukotrieens in hypoxic pulmonary vasoconstriction in rats.

The role of leukotrienes in the mechanism of hypoxic pulmonary vasoconstriction (HPV) is controversial. To determine whether leukotriene C4 (LTC4) was produced during HPV, LTC4 levels were measured in individual samples of lung tissue, lung bronchoalveolar lavage fluid (BALF), and blood perfusate in isolated perfused lungs ventilated with normoxic or hypoxic gas mixtures. HPV was not associated with increased LTC4 in lung tissue or increased LTE4 in blood perfusate. Consistent with previous studies demonstrating elevated levels of LTC4 in pooled BALF fluid from hypoxic lungs, individual lung BALF samples demonstrated an elevation of LTC4 during hypoxia. However, the process of lung lavage alone stimulated eicosanoid production, with LTC4, 6-ketoprostaglandin F1 alpha, and thromboxane B2 levels being higher in lavaged compared to non-lavaged lungs. In lungs to which the lipoxygenase inhibitor AA 861 was added to the perfusate, a reduction in the lung tissue LTC4 levels was observed without any or only a slight reduction in HPV. To evaluate the physiological effects of LTC4 in the airways, exogenous LTC4 (1-1,000 ng) was added to the airways of both blood- and physiological salt solution-perfused lungs without any effect on the pulmonary artery pressure or a response to hypoxia. These results do not support the hypothesis that leukotrienes mediate HPV in the rat.

Animals↗