Carlo Vicentini

Epidermal growth factor modulates prostate cancer cell invasiveness regulating urokinase-type plasminogen activator activity. EGF-receptor inhibition may prevent tumor cell dissemination.

Urokinase-type plasminogen activator receptor (uPAR) and Epidermal Growth Factor Receptor (EGFR) are ubiquitous receptors involved in the control of a variety of cellular processes frequently found altered in cancer cells. The EGFR has been recently described to play a transduction role of uPAR stimuli, mediating uPA-induced proliferation in highly malignant cells that overexpress uPAR. We compared the uPA production, the presence of uPAR, AR, EGFR and Her2 with the chemotaxis and the Matrigel invasion in ten human PCa cell lines and observed that: (1) the levels of Her2, but not of EGFR, as well as the uPA secretion, cell motility and Matrigel invasion were statistically higher in AR negative than in AR positive PCa cells; (2) the uPA secretion and uPA Rexpression were positively related to Matrigel invasion; (3) the EGF was able to stimulate chemotaxis and Matrigel invasion in a dose-dependent manner; (4) the EGF-induced cell migration was statistically higher inAR negative than in AR positive cells with a similar increase with respect to basal value (about 2.6 fold); (5) the Matrigel invasion was statistically higher in AR negative than in AR positive PCa cells also if the increment of Matrigel invasion after EGF treatment was statistically higher in AR positive respect to AR negative cells; (6) the EGF induced uPA secretion and its membrane uptake through the increment of uPAR; and (7) these effects were blocked by EGFR/Her2 tyrosine kinase inhibitors with IC(50) lower than those needed to inhibit cell proliferation and required PI3K/Akt, MAPK and PI-PLC activities as verified by inhibition experiments. These enzymatic activities were regulated in different manners in PTEN positive and negative cells. In fact, the inhibition of PI3K blocked the EGF-induced invasiveness in PTEN positive cells but not in PTEN negative cells, in which PI3K activity was not influenced by EGFR/Her2 activation, whereas the inhibition of MAPK was able to block the invasive phenomena in both cell types. Taken together, our data suggest that the blockade of EGFR could attenuate the invasive potential of PCa cells. In addition, considering that the EGFR expression is related to higher Gleason grade of PCa and that EGFR levels are increased after anti androgenic therapy, this therapeutic approach could slow down the metastasis formation which represents the most dramatic event of PCa progression.

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.

Azacitidine improves antitumor effects of docetaxel and cisplatin in aggressive prostate cancer models

One of the major obstacles in the treatment of hormone-refractory prostate cancer (HRPC) is the development of chemoresistant tumors. The aim of this study is to evaluate the role of azacitidine as chemosensitizing agent in association with docetaxel (DTX) and cisplatin using two models of aggressive prostate cancer, the 22rv1, and PC3 cell lines. Azacitidine shows antiproliferative effects associated with increased proportion of cells in G0/G1 and evident apoptosis in 22rv1 cells and increased proportion of cells in G2/M phase with the absence of acute cell killing in PC3 cells. In vivo, azacitidine (0.8 mg/kg i.p.) reduced tumor proliferation and induced apoptosis in both xenografts upmodulating the expression of p16INKA, Bax, Bak, p21/WAF1, and p27/KIP1, and inhibiting the activation of Akt activity and the expression of cyclin D1, Bcl-2, and Bcl-XL. In vitro treatments with azacitidine lead to upregulation of cleaved caspase 3 and PARP. BCl2 antagonists, such as HA-14-1, enhanced the effects of azacitidine in these two prostate cancer models. In addition, azacitidine showed synergistic effects with both DTX and cisplatin. In vivo this agent caused tumor growth delay without complete regression in xenograft systems. Azacitidine sensitized PC3 and 22rv1 xenografts to DTX and cisplatin treatments. These combinations were also tolerable in mice and superior to either agent alone. As DTX is the standard first-line chemotherapy for HRPC, the development of DTX-based combination therapies is of great interest in this disease stage. Our results provide a rationale for clinical trials on combination treatments with azacitidine in patients with hormone-refractory and chemoresistant prostate tumors.

Sexual function in women with urinary incontinence treated by pelvic floor transvaginal electrical stimulation.

INTRODUCTION Womens sexual dysfunctions (WSD) have been commonly associated with urinary incontinence (UI). Women with UI and who scored low on the Female Sexual Function Index (FSFI) showed an improvement in urinary leakage and also in their sexual life following treatment by transvaginal electrical stimulation (TES). AIMS To determine the effects of TES in 37 women complaining of UI, of whom 23 also had WSD, and to compare the FSFI scores of women with UI and 43 women not affected by UI who underwent routine urologic evaluation. METHODS Thirty-seven women complaining of UI were evaluated by voiding diary and with FSFI before and after 3 months of TES. All had a urogynecologic evaluation and urodynamic study. MAIN OUTCOME MEASURES In the voiding diary the women reported the types of liquid they ingested, urinary frequency, and episodes of urgency and urine leakage. The domain scores of the FSFI, including desire, arousal, lubrication, orgasm, satisfaction, and pain, were calculated. TES was conducted for 15-30 minutes, twice a week for 3 months, using biphasic intermittent current with a frequency of 50 Hz for stress UI (SUI) and 20 Hz for urge UI (UUI), and the most tolerable intensity of stimulation. RESULTS After TES, only two of the 10 women with UUI experienced a few leakage incidents; patients with SUI were completely dry during TES; and only three reported a few episodes of UI during intense activities. The five patients with mixed UI improved mainly as regards urgency. The FSFI scores of patients complaining of UI showed significantly lower desire and sexual satisfaction, and higher sexual pain than controls. After 3 months, the 23 women affected by WSD, of the 37 participants with UI, reported a remarkable improvement in their sexual life. CONCLUSIONS TES was found to be a safe and effective therapy for selected patients affected by mild to moderate UI. Because women with UI also complain of WSD compared with the general female population, an investigation of female sexuality is suggested for these patients.

Additive antitumor effects of the epidermal growth factor receptor tyrosine kinase inhibitor, gefitinib (Iressa), and the nonsteroidal antiandrogen, bicalutamide (Casodex), in prostate cancer cells in vitro.

Progression from an androgen‐dependent to an androgen‐independent state often occurs in patients with prostate cancer (PCa) who undergo hormonal therapy. We have investigated whether inhibition of the epidermal growth factor receptor (EGFR) signaling pathway affects the antitumor effect of a nonsteroidal antiandrogen. Gefitinib (Iressa), an EGFR tyrosine kinase inhibitor, and bicalutamide (Casodex), a nonsteroidal antiandrogen [androgen receptor (AR) antagonist], were administered alone and in combination to AR‐positive human PCa cell lines. FACS analysis showed lower EGFR expression levels on AR‐positive cells (LNCaP, CWR22, CWR22R 2152 and AR‐transfected DU145 cell lines) compared with AR‐negative cells (DU145, PC3 and TSU‐Pr1). Moreover, in AR‐transfected DU145 cells, chronic treatment with bicalutamide increased EGFR expression to levels similar to androgen‐independent DU145 cells. All AR‐positive PCa cell lines were sensitive to gefitinib (IC50 = 0.1–0.6 μM), whereas higher concentrations of bicalutamide were needed to reduce AR‐positive PCa cell line proliferation (IC50 = 0.8–2.0 μM). Low doses of gefitinib increased the antitumor effects of bicalutamide by strongly reducing the IC50 of bicalutamide (approximately 10‐fold). Similarly, bicalutamide increased the antiproliferative effects of gefitinib by reducing the IC50 of gefitinib (approximately 5‐fold). Taken together, our data suggest that in androgen‐dependent cell lines, addition of gefitinib in combination with bicalutamide results in concurrent dual inhibition of AR and EGFR/HER2 pathways. This causes a significant delay in the onset of EGFR‐driven androgen independence.

Body mass index, urinary incontinence, and female sexual dysfunction: how they affect female postmenopausal health.

Objective: The aim of this study was to evaluate the relationship between body mass index (BMI) and female sexual dysfunction (FSD) among perimenopausal and postmenopausal women with urinary incontinence (UI). Methods: From 2005 to 2008, we enrolled 208 consecutive women affected by UI; all underwent a comprehensive history including two validated questionnaires, physical examination, and urodynamic evaluation. Based on BMI, participants were grouped into normal weight, overweight, and obese. Results: A total of 158 participants completed both questionnaires (76% response rate); 41 (26%) were normal weight, 73 (46%) were overweight, and 44 (28%) were obese. The increasing Urogenital Distress Inventory score had a direct correlation with age (P < 0.01), year of menopause onset (P < 0.05), and BMI (P < 0.01). FSD was diagnosed in 97 women (61%): 31 (32%) with hypoactive sexual desire, 20 (21%) with sexual arousal disorder, 7 (7%) with orgasmic deficiency, and 39 (40%) with sexual pain disorder. BMI greater than 30 kg/m2 was independently associated with an increased risk of FSD (odds ratio [OR], 2.02) and UI (OR, 2.03). With adjustment for BMI, the OR for FSD was 1.22 for overweight women and 1.56 for obese women, with respect to healthy participants. The total Female Sexual Function Index score correlated with BMI (r = −0.82, P = 0.0001); in particular, arousal (r = −0.82), orgasm (r = −0.72), lubrication (r = −0.61), and satisfaction (r = −0.63, all P < 0.001) showed an inverse correlation with BMI, whereas desire and pain did not. Conclusions: Increased BMI early in menopause represents a risk both for UI and for sexual dysfunction. Weight control has an essential role in postmenopause and should be considered early in perimenopause to safeguard female quality of life as well as to prevent or improve UI and female sexual dysfunction symptoms.

Oxidative Stress in Benign Prostatic Hyperplasia and Prostate Cancer

Objective: The aim was to verify whether oxidative stress could represent a common key factor of benign prostatic hyperplasia (BPH) and prostate cancer (PCa). Subjects and Methods: 15 patients affected by BPH, 15 with PCa and 15 controls were enrolled. Blood samples were withdrawn systemically and locally during radical retropubic prostatectomy in patients with PCa and during transvesical retropubic adenomectomy in patients diagnosed with BPH. Plasma oxidized low-density lipoprotein, peroxides, and total equivalent antioxidant capacity (TEAC) including plasma superoxide dismutase (SOD) determination were analyzed as oxidative markers. Results: With respect to the control group, high plasma peroxides and decreased TEAC levels were measured in patients affected by both PCa and BPH. Plasma peroxides were significantly higher in patients with PCa with respect to BPH. A positive correlation was found between peroxides and TEAC values in samples withdrawn locally in patients affected by PCa. An inverse correlation between peroxides and TEAC was observed in patients with BPH. No statistically significant modifications were observed as concerns SOD activity and LDL oxidability. Conclusions: Our findings confirm a significant unbalance of redox status in patients affected by BPH and PCa, and suggest a potential involvement of oxidative stress as a determinant in the pathogenesis of these diseases.

Akt down-modulation induces apoptosis of human prostate cancer cells and synergizes with EGFR tyrosine kinase inhibitors

PTEN is a well‐characterized tumor suppressor that negatively regulates cell growth and survival through the modulation of PI3K/Akt pathway.

Plasmakinetic bipolar versus monopolar transurethral resection of non‐muscle invasive bladder cancer: A single center randomized controlled trial

To compare the safety and the efficacy of plasmakinetic bipolar resectoscope versus conventional monopolar in the transurethral resection of primary non‐muscle invasive bladder cancer.

The Role of Left Superior Parietal Lobe in Male Sexual Behavior: Dynamics of Distinct Components Revealed by fMRI

INTRODUCTION Despite the interest for the brain correlates of male sexual arousal, few studies investigated neural mechanisms underlying psychogenic erectile dysfunction (ED). Although these studies showed several brain regions active in ED patients during visual erotic stimulation, the dynamics of inhibition of sexual response is still unclear. AIM This study investigated the dynamics of brain regions involved in the psychogenic ED. METHODS Functional magnetic resonance imaging (fMRI) and simultaneous penile tumescence (PT) were used to study brain activity evoked in 17 outpatients with psychogenic ED and 19 healthy controls during visual erotic stimulation. Patterns of brain activation related to different phases of sexual response in the two groups were compared. MAIN OUTCOME MEASURES Simultaneous recording of blood oxygen level-dependent fMRI responses and PT during visual erotic stimulation. RESULTS During visual erotic stimuli, a larger activation was observed for the patient group in the left superior parietal lobe, ventromedial prefrontal cortex, and posterior cingulate cortex, whereas the control group showed larger activation in the right middle insula and dorsal anterior cingulate cortex and hippocampus. Moreover, the left superior parietal lobe showed a larger activation in patients than controls especially during the later stage of sexual response. CONCLUSION Our results suggest that, among regions more active in patient group, the left superior parietal lobe plays a crucial role in inhibition of sexual response. Previous studies showed that left superior parietal lobe is involved in monitoring of internal body representation. The larger activation of this region in patients during later stages of sexual response suggests a high monitoring of the internal body representation, possibly affecting the behavioral response. These findings provide insight on brain mechanisms involved in psychogenic ED.