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Biomedical subjects

M Bologna

Publications and source records attributed to M Bologna.

At least 37 records · Page 2Linked to original sources
PubMed1999

Osteoblasts modulate secretion of urokinase-type plasminogen activator (uPA) and matrix metalloproteinase-9 (MMP-9) in human prostate cancer cells promoting migration and matrigel invasion.

Prostate carcinoma (PRCA) cells metastasize to the skeleton with high frequency. Bone stores growth regulatory factors, which are released in active form during bone remodeling. We propose that bone cell-derived growth factors may induce the development of PRCA bone metastasis by recruiting tumor cells and increasing their proliferation in the bone microenvironment. Serum-free conditioned medium harvested from osteoblast cultures (OB CM) stimulated the in vitro chemotaxis of PRCA cells and invasion of a reconstituted basement membrane (Matrigel), suggesting enhanced invasive activity. Preosteoblastic cell CMs were less effective than CMs obtained from mature OB. CMs harvested from differentiated osteoblast cultures capable of matrix mineralization were more active compared to CMs from proliferating osteoblasts. OB CMs stimulated secretion of urokinase (uPA) and matrix metalloproteinase-9 (MMP-9). Inhibition of these matrix-degrading proteases by neutralizing antibodies and/or by inhibitors of their catalytic activity reduced Matrigel invasion. Secretion of uPA and activation of MMP-9 were most prominent by differentiated OB CMs with respect to poorly differentiated cells in vitro. These results are in agreement with several in vivo studies and indicate that factors produced during osteogenesis by bone cells stimulate PRCA cell chemotaxis and matrix proteases expression, thus representing potential targets for alternative therapies deterring the progression of PRCA metastasis to bone.

Cell Movement↗
PubMed1998

In vitro regulation of pericellular proteolysis in prostatic tumor cells treated with bombesin.

Bombesin is a potent inducer of signal trasduction pathways involved in the proliferation and invasion of androgen-insensitive prostatic tumor cells. This study examines the bombesin-mediated modulation of pericellular proteolysis, monitoring cell capability to migrate and invade basement membranes, using a chemo-invasion assay and analyzing protease production. The results suggest that bombesin could modulate the invasive potential of prostatic cell lines regulating secretion and cell-surface uptake of uPA and MMP-9 activation. In fact, in PC3 and DU145 cells but not in LNCaP cells, urokinase-type plasminogen activator (uPA) and plasminogen activator inhibitor-1 (PAI-1) are induced by bombesin treatment. Bombesin also stimulates cell proliferation and this effect can be inhibited blocking uPA by antibodies and/or uPA inhibitor p-aminobenzamidine. Moreover, HMW-uPA induces cell proliferation in LNCaP cells, which do not produce uPA in the basal conditions, while PC3 and DU145 cell growth is supported by autocrine production of uPA. The increment of uPA activity on the external plasma membrane causes an increased pericellular plasmin activation. This effect is inhibited by antibodies against uPA and by p-aminobenzamidine. Similarly to EGF, bombesin stimulates secretion and activation of MMP-9 and TIMP-1 production. MMP-9 activation can be also obtained by HMW-uPA treatment, suggesting that plasma-membrane-bound uPA can start a proteolytic cascade involving MMP-9. Therefore, in in vitro assays, bombesin is able to modulate pericellular proteolysis and cell proliferation, differently distributing and activating proteolytic activities. This effect can be related to the "non-random" degradation of the extracellular matrix in which membrane uPA-uPAreceptor complexes could start bombesin-induced directional protein degradation during metastatic spread.

Antibodies↗
PubMed1998

Plasminogen activator system modulates invasive capacity and proliferation in prostatic tumor cells.

The malignant phenotype of prostatic tumor cells correlates with the expression of both uPA and its cell-membrane receptor (uPAR); however, there is little information concerning the role of cell-bound uPA in matrix degradation and invasion. Our results suggest that cell-associated uPA plays a key role in regulating the amount of plasmin present at the surface of prostatic carcinoma (PRCA) cells and show that differential production of uPA corresponds with the capacity to bind and activate plasminogen. In addition, we provide direct evidence that both uPA secretion and the presence of uPA-uPAR complexes characterize the invasive phenotype of PRCA cells and suggest the existence of several pathways by which tumor cells acquire plasmin activity. LNCaP cells (which do not produce uPA but express uPAR) may activate plasmin through exogenous uPA. In vivo, the source of uPA may be infiltrating macrophages and/or fibroblasts as observed in several other systems. PAI-1 accumulation in the conditioned medium (CM) limits plasmin action to the pericellular microenvironment. Our results indicate that MMP-9 and MMP-2 are also activated by plasmin generated by cell-bound but not by soluble, extracellular uPA. Plasmin activation and triggering of the proteolytic cascade involved in Matrigel invasion is blocked by antibodies against uPA (especially by anti- A-chain of uPA which interacts with uPAR) and by PA inhibitors such as p-aminobenzamidine which may regulate levels of cell-bound uPA. uPA may also regulate growth in PRCA cells. Indeed, antibodies against uPA A-chain (and also p-aminobenzamidine treatment) interfere with the ATF domain and inhibit cell growth in uPA-producing PC3 and DU145 prostate cancer cell lines, whereas exogenous uPA (HMW-uPA with ATF) induces growth of LNCaP prostate tumor cell line. These data support the hypothesis that in prostatic cancer patients at risk of progression, uPA/plasmin blockade may be of therapeutic value by blocking both growth of the primary tumor and dissemination of metastatic cells.

Benzamidines↗
PubMed1997

Human prostatic tumor cells in culture produce growth and differentiation factors active on osteoblasts: a new biological and clinical parameter for prostatic carcinoma.

Prostate cancer (PRCA) cells metastasize to bone with high frequency, inducing typical osteosclerotic lesions. To establish if local stimuli on the bone tissue may derive from metastatic colonies of prostatic origin, we evaluated the biologic activities secreted by human prostatic epithelium and effective on osteoblast-like cells in vitro. Supernatant from short-term tissue cultures of human prostatic tissue samples obtained from PRCA (35 cases) and benign prostatic hyperplasia (BPH, 12 cases) patients were applied to three models of cells with osteoblastic phenotype: two normal [rabbit osteoblasts (OB) and rat periosteal cells (PO)] and one transformed (human osteosarcoma cell line, MG63). Proliferative activity was monitored through enzymatic reduction of tetrazolium salts and expressed as relative mitogenic activities (RMA). Analysis of proliferation and alkaline phosphatase (ALP) activity, a marker of osteoblast function, demonstrates that conditioned media (CM) from PRCA cultures stimulate both growth and activity of osteoblast-like cells to a greater extent compared to CM from BPH. Furthermore, cell growth and activity of osteoblast-like cells are progressively increased by CM derived from patients with stage B (tumor confined within the prostate capsule), stage C (locally invasive tumor), and stage D (invasive tumor with distant metastasis) disease. One of the mechanisms potentially underlying the CM-stimulated effects on bone cells is associated with the urokinase (uPA) enzyme route, whose release progressively increases with the stage of disease. However, antibodies against uPA and p-aminobenzamidine (a low molecular weight urokinase inhibitor) treatment, which both inhibit the proliferative and differentiative effects induced by exogenous urokinase, partially slow down the effects of CM from PRCA tissue cultures, suggesting that additional factors are secreted by prostatic tumor cells in vitro. In conclusion, we show that the mitogenic and differentiative activities for osteoblasts produced by prostatic tumor cells in short-term tissue cultures are related to PRCA stage and may predict the behavior of skeletal metastases in single cases of tumor. In addition, the culture methods used may represent a valid model to study prostatic and bone cellular interactions, which may indicate new therapeutic approaches in metastatic prostate tumors.

Alkaline Phosphatase↗
PubMed1996

Increased matrix metalloproteinase-9 secretion in short-term tissue cultures of prostatic tumor cells.

We have examined the expression of 2 tumor-associated metalloproteinases, MMP-2 and MMP-9, in 48 primary cultures of prostatic carcinoma (PRCA) and 33 cultures of benign prostatic hyperplasia (BPH). PRCA cultures secrete significantly more MMP-9 than their benign counterparts. Secreted MMP-2 did not differ significantly in cultures but was lower in PRCA cultures. Two cultures of benign origin exhibited high MMP-9 secretion and growth patterns consistent with a malignancy. Both cases were followed and successively re-evaluated histologically and rediagnosed as organ-confined PRCA. MMP expression in culture may be of predictive value in the identification of incidental PRCA. MMP-9 secretion and its ratio with MMP-2 were highest in epithelial cultures from invasive, metastatic tumors when compared both to disease confined to prostate gland and to locally extensive disease. MMP-9 secretion was greatest also in cultures derived from tissues of high Gleason histological grade. Active MMP-9 species were detected in 15 cultures (31%) of PRCA. Active MMP-2 species were observed in cultures of both BPH and PRCA origin in almost the same amounts. Although average levels were not significantly different, as a ratio to proform species, a significant elevation was observed in cultures of PRCA origin. We propose, therefore, that an elevated expression of MMP-9 and a high ratio of MMP-9 to MMP-2 in short-term prostate epithelial cultures is of potential diagnostic and prognostic significance.

Diagnosis, Differential↗
PubMed1995

Finasteride dose-dependently reduces the proliferation rate of the LnCap human prostatic cancer cell line in vitro.

OBJECTIVES: To assess the effects of finasteride, a 5-alpha-reductase inhibitor, and of classic antiandrogens on the growth rate of the LnCap human prostate carcinoma cell line, derived from a primary and well-differentiated neoplasm. METHODS: Cell proliferation experiments in vitro with and without the antiandrogens cyproterone acetate, hydroxyflutamide, and finasteride in the 0.0001 to 10.0 microM range. RESULTS: The growth rate of the LnCap cell line can be dose-dependently inhibited by 5-alpha-reductase inhibition (finasteride) and by antiandrogens (cyproterone acetate and hydroxyflutamide) in vitro, in defined conditions. CONCLUSIONS: Besides other human prostate cell lines derived from metastatic sites (PC3, DU145), also in the LnCap cell line an autonomous androgen-dependent mechanism of growth stimulation can be hypothesized, since testosterone and dihydrotestosterone are unable to stimulate the cell proliferation rate at the same molar concentrations. The clinical implications of these results in prostate cancer therapy and the possible future use of these molecules in the prevention of cancer incidence are discussed.

Androgen Antagonists↗
PubMed1995

Improved tissue culture method for the study of prostatic carcinoma: a significant diagnostic tool.

A new in vitro method for early detection of prostatic cancer is described. The results of tissue culture of prostatic epithelial cells harvested during digital rectal examination were compared with patient-matched biopsy findings in a double-blind study. An arbitrary combination of five culture parameters (displayed in vitro by cultured prostatic cells and related to neoplastic and differentiation features of the sample [growth capacity, proportion of epithelium and morphologic atypia of overlapping cells, multinucleated cells and vacuolized cells]), was created after two weeks of culture; this combination, referred to as the Global Tissue Culture Score (GTCS), ranged from 0 (negative result or benign behavior) to 15 (highly abnormal neoplastic behavior). The GTCS was significantly higher in biopsy-proven prostatic carcinoma than in other prostatic diseases. The combination of GTCS and patient age was highly sensitive and specific for a diagnosis of prostatic carcinoma, according to the biopsy results. Our findings indicate that the early diagnosis of prostatic carcinoma can be obtained by a tissue culture method which allows in vitro growth of epithelial cells harvested through digital rectal prostatic massage. This method is easy to perform, is non-invasive, is able to discriminate between biologically aggressive and non-aggressive neoplasms, and may be useful for mass screening of prostatic carcinoma in conjunction with digital rectal exam. Early diagnosis of prostatic carcinoma may maximize the potential curative value of radical prostatectomy or other forms of therapy for this highly prevalent cancer.

Adult↗
PubMed1995

Plasminogen activator activities in short-term tissue cultures of benign prostatic hyperplasia and prostatic carcinoma.

The high prevalence of prostatic carcinoma (PRCA) and the limited therapeutic possibilities provide a strong stimulus for exploring new approaches in experimental research that ultimately may lead to improved therapy. Indeed, methods for assessing carcinoma prognosis, such as clinical staging (clinical examination, ultrasound, and plasmatic levels of prostatic acid phosphatase and prostate specific antigen) and histopathological grading according to the Gleason score, usually fail to provide consistent predictive information regarding the clinical outcome of single tumors. Increased plasminogen activator (PA) activities have been associated with high-grade malignancies and with the potential for invasion/metastasis in many tumors. Urokinase-type plasminogen activator (uPA) is present in prostatic secretion, and an increased uPA activity has been noted in human prostatic cell lines with metastatic behavior. Unfortunately, any study of uPA production or gene regulation in primary tumors is complicated by the inherent mixture of host stromal cells, infiltrating macrophages, and subpopulations of tumor cells that may have variable metastatic capacity and ability to synthesize uPA. In short-term tissue culture of prostatic samples, it is possible to grow in vitro cancer prostatic epithelial cells and thus exclude the presence of contaminant cells. We have shown elsewhere that the levels of a type IV collagenase, 92-kDa matrix metalloproteinase, a protease involved in tumor progression and invasion, are increased in PRCA primary cell cultures if compared with benign prostatic hyperplasia (BPH) cell cultures (C. Festuccia et al., manuscript in preparation). Activation of matrix metalloproteinases also can be correlated with uPA expression; therefore we studied the expression of uPA in serum-free culture media of primary cultures of PRCA or BPH tissue samples.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗
PubMed1993

Early diagnosis of prostatic carcinoma may be achieved through in vitro culture of tumor cells harvested by prostatic massage.

As a new method for early diagnosis of prostatic carcinoma we succeeded in growing in vitro the epithelial cells which can be collected from prostatic fluid after rectal prostatic massage. We report here the updated and statistically analyzed series of data (174 patients) on this new approach, which allows all the harvested cells to express their biological features. The method reaches a sensitivity of 72-86% and a specificity of 88-100%. This noninvasive test, which is also suitable for mass screening, may be very useful for an early diagnosis of the neoplasm.

Adult↗
PubMed1993

Neuroendocrine epithelial cells of the main pancreatic ducts and ampulla.

In this study we investigated by immunocytochemistry (peroxidase-conjugated extravidin-biotin system) the existence of neuroendocrine cells in the main ducts draining the exocrine pancreas and in the ampulla of Vater of six human specimens obtained at surgery (duodenocephalopancreasectomy). Using a panel of primary antibodies for Chromogranin A, gastrin, CCK, somatostatin and serotonin (5HT), we visualized scattered neuroendocrine cells in the epithelium of Wirsung duct and in the ampulla. All were positive for Chromogranin A and the majority were also positive for serotonin (5HT), while gastrin cells were seldom observed. Interestingly, their number was far lower than in the adjacent duodenal mucosa. We believe that these neuroendocrine epithelial cells may have specific physiological roles such as epithelial trophism and differentiation.

Aged↗
PubMed1992

Simultaneous determination of simvastatin and its hydroxy acid form in human plasma by high-performance liquid chromatography with UV detection.

Simvastatin (SV), an analogue of lovastatin, is the lactone form of 1',2',6',7',8',8a'-hexahydro-3,5-dihydroxy-2',6'-dimethyl-8'(2'',2''-di met hyl-1''-oxobutoxy)-1'-naphthalene-heptanoic acid (SVA) which lowers plasma cholesterol by inhibiting 3-hydroxy-3-methylglutaryl-CoA reductase. A fast, simple and accurate method for determining SV and SVA concentrations in human plasma has been developed and validated for use in the analysis of plasma samples from patients and healthy volunteers. This method involves an extraction procedure using a mixture of acetonitrile-water and reversed-phase high-performance liquid chromatography with UV detection. The procedure was linear from 20 to 1000 ng ml-1 for SV and from 25 to 1000 ng ml-1 for SVA, respectively. The method was accurate with relative errors of 5.0, 2.1 and 3.2% for human plasma controls containing 50, 250 and 500 ng ml-1 of SV, respectively. The corresponding precision was 2.3, 1.8 and 1.0% (RSD%). Similarly, relative standard deviations less than 2.3% and relative errors of less than 5.2% were obtained from human plasma controls containing SVA at identical concentrations. The method is suitable for pharmacology and pharmacokinetic studies of simvastatin.

Blood Proteins↗
PubMed1992

Sex-role change, anomie and female suicide: a test of alternative Durkheimian explanations.

Durkheim's influential book, Suicide, provides general (economic) anomie, conjugal anomie, and sex-role convergence explanations of changes in suicide rates under conditions of social change. We used trend analyses of American suicide rates and female/male suicide ratios from 1950 to 1984 and the regression of the ratios on female educational attainment, white female labor force participation, white fertility rates, and divorce rates to examine these explanations. The general anomie explanation of female suicide trends is supported for the middle-aged (30 to 54 years) but not for the young (10 to 30 years) or the elderly (55 to 74 years). The conjugal anomie proposition is at best supported for age groups between 15 and 44 when general anomie is not pronounced. The role convergence explanation is rejected for all 13 5-year-age-groups. General anomie may not be a viable explanation of suicide trends for groups actively supporting relevant social changes or not yet tradition-bound or for groups whose retirement status reduces the importance of some social changes.

Adult↗
PubMed1991

Establishment and characterization of a human prostatic carcinoma cell strain obtained from exfoliated tumor cells after prostatic massage.

During a control campaign connected to our main program on early diagnosis of prostatic carcinoma through tissue culture of prostatic fluid samples obtained after prostatic massage (M. Bologna et al., Eur. Urol., 14, 474-476, 1988), we isolated and characterized a human prostatic carcinoma cell strain from a 58-year-old patient with a grade III prostatic carcinoma. The epithelial cell strain, named PMU-23, has been passaged in vitro for 31 subculture cycles during a period of approximately 8 months, after which cell proliferation slowed down irreversibly. The isolation of this cell strain constitutes a renewed confirmation of the validity of our method for the early diagnosis of prostatic carcinoma and demonstrates some intermediate features in the progression of prostatic tumors. In addition, the study of limited-lifespan tumor cell strains in culture may extend the knowledge on prostatic cell biology, particularly toward the identification of intermediate steps of tumor progression, for a better approach of tumor therapy and prevention of metastatic spread.

Cell Division↗
PubMed1989

Bombesin stimulates growth of human prostatic cancer cells in vitro.

Cell proliferation of the human prostatic carcinoma cell line PC3 and of the epithelial cell strain PMU 23 derived from a primary culture of a stage III prostatic carcinoma was enhanced dose dependently by adding 0.1 nM to 10.0 nM bombesin (BMBS) to the culture medium. The growth stimulation was specifically inhibited by antibodies versus Gastrin Releasing Peptide (GRP) crossreacting with BMBS. Presence of BMBS-positive neuroendocrine cells in human prostate and measurable amounts of BMBS-like peptides in prostatic fluid were reported previously. In a binding assay using 125I-GRP, it was possible to demonstrate the presence of saturable specific receptors on PC3 cells, numerically comparable with those measured on small cell lung cancer cell lines. By immunofluorescence, however, no BMBS immunoreactivity on PC3 cells could be demonstrated. These observations suggest that BMBS plays a role in prostatic epithelium growth and that prostatic carcinoma may have an autocrine or paracrine proliferation stimulus within the gland microenvironment.

Bombesin↗