Barbara Jackson

Proteomic analysis of SEG-1 human Barrett's-associated esophageal adenocarcinoma cells treated with keyhole limpet hemocyanin

Keyhole limpet hemocyanin (KLH) is an immune stimulant derived from a circulating glycoprotein of the marine mollusk Megathura crenulata. We previously reported that KLH inhibited the growth of human Barrett’s-associated esophageal adenocarcinoma in vitro via apoptotic and nonapoptotic mechanisms. We hypothesize that KLH reduces the growth of Barrett’s cancer cells by altering protein expression profiles. A cell line (SEG-1) derived from Barrett’s-associated adenocarcinomas of the distal esophagus was selected. Cells were administered KLH (500 µg/ml) or vehicle. After 24 hours, cytosolic fractions were separated through two-dimensional gel electrophoresis. Statistical analysis was performed with Evolution Pro software to identify spots that were differentially expressed between the KLH and control groups. Proteins displaying a twofold or greater change in expression levels were selected for identification. In a total of 420 spots, 31 were differentially expressed between the KLH and control groups. In all, 12 were upregulated and 19 were downregulated. Of the 31, 17 were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Proteomic evaluation shows downregulation of proteins associated with metabolic processes (glycolysis, protein synthesis). KLH also induced proteins indicative of oxidative stress (heat shock 70 family and UDP-glucose 6-dydrogenase). Our results indicate that growth arrest by KLH is accompanied by a cellular stress response and attenuation of metabolic processes. The use of KLH as adjuvant or topical therapy for Barrett’s adenocarcinoma provides a promising development in the treatment of this disease.

Effect of chronic stress on running wheel activity in mice

Acute and chronic stress have been reported to have differing effects on physical activity in rodents, but no study has examined a chronic stress protocol that incorporates stressors often experienced by rodents throughout a day. To examine this, the effects of the Unpredictable Chronic Mild Stress (UCMS) protocol on voluntary running wheel activity at multiple time points, and/or in response to acute removal of chronic stress was determined. Twenty male Balb/c mice were given access and accustomed to running wheels for 4 weeks, after which they were randomized into 2 groups; exercise (EX, n = 10) and exercise with chronic stress using a modified UCMS protocol for 7 hours/day (8:00 a.m.-3:00p.m.), 5 days/week for 8 weeks (EXS, n = 10). All mice were given access to running wheels from approximately 3:30 p.m. to 7:30 a.m. during the weekday, however during weekends mice had full-time access to running wheels (a time period of no stress for the EXS group). Daily wheel running distance and time were recorded. The average running distance, running time, and work each weekday was significantly lower in EXS compared to EX mice, however, the largest effect was seen during week one. Voluntary wheel running deceased in all mice with increasing age; the pattern of decline appeared to be similar between groups. During the weekend (when no stress was applied), EXS maintained higher distance compared to EX, as well as higher daily distance, time, and work compared to their weekday values. These results indicate that mild chronic stress reduces total spontaneous wheel running in mice during the first week of the daily stress induction and maintains this reduced level for up to 8 consecutive weeks. However, following five days of UCMS, voluntary running wheel activity rebounds within 2–3 days.

In Vitro Regulation of Cell Growth and Angiogenesis by Inositol Hexaphosphate in Bladder Cancer

Background: Inositol Hexaphosphate (IP6) is a naturally occurring polyphosphorylated carbohydrate that is found in food sources high in fiber content. We hypothesized that IP6 would inhibit the cell growth rate of bladder cancer in vitro. Methods: T24 and TCCSUP bladder cancer cell lines were treated with titrating doses of IP6 (0.3, 0.6 and 0.9 mM/well). Cell viability and vascular endothelial growth factor levels were measured. Results: Significant reductions (p < 0.001) in cellular growth were noted in both cell lines at all doses and time points tested, with the exception of 0.3 mM IP6 at 24 hours in the T24 cell line. The percent inhibition of vascular endothelial growth factor was significantly higher than that observed in the TCCSUP cell line at 48 and 72 hours with 0.3 mM IP6 (p < 0.001). The T24 cells exhibited the same level of inhibition at 24 and 48 hours with 0.6 mM dose of IP6 and at 72 hours with the 0.3 mM dose (p < 0.001). Conclusions: In vitro treatment of bladder cancer with the common dietary polyphosphorylated carbohydrate IP6 significantly decreased cellular growth by anti-angiogenic mechanisms. We feel that this data warrants further investigation and consideration for initiation of clinical trials to evaluate the safety and clinical utility of this agent.

Inositol Hexaphosphate (IP6): Modulation of Cell Cycle and Proliferation of Bladder Cancer in vitro

Introduction: We hypothesized that inositol hexaphosphate (IP6) would modulate cell cycle and cellular growth in bladder cancer. Materials and Methods: Bladder cancer cell lines were treated with 2.5 or 4.5 mM/well IP6. Cell viability was measured by MTT at 24 and 48 h. Cell cycle analysis was measured by DNA staining and quantified by FACS analysis. Results: Cell growth was inhibited in the TCCSUP, HTB9 and T24 cells (p < 0.001) with the exception of the HTB9 cells at 24 h. G1 phase fractions increased in the TCCSUP and HTB9 cells, while S phase fraction was decreased (p < 0.001). G2 phase was decreased in the TCCSUP cells (p < 0.001). G1 phase fraction was decreased in the T24 line with 4.5 mMIP6 (p < 0.001), while a significant increase in the S phase was observed in the T24 cells (p < 0.001). Conclusions: These results indicate that IP6 is potentially a highly effective treatment for carcinoma of the bladder.

PD44-07 FEMALE SEXUAL DYSFUNCTION: A WEST VIRGINIA UNIVERSITY CLINICAL EXPERIENCE

INTRODUCTION AND OBJECTIVES: Female lower urinary tract symptoms (LUTS) affect quality of life and sexual activity. This study aimed to evaluate the influences of LUTS on sexual well-being in Japanese women, as little is known on this topic. METHODS: We investigated 514 women recruited between August 6 and August 17, 2007, from the outpatient departments (except the departments of pediatrics, psychiatry and ophthalmology) at our hospital, regardless of the reason for visiting. All participants were asked to answer a standardized self-reported questionnaire. Using the International Consultation on Incontinence Questionnaire-Short Form (ICIQ-SF) and the overactive bladder symptom score (OABSS), we evaluated urinary symptoms, including stress urinary incontinence, urgency, day time frequency, and nocturia. To assess satisfaction with sexual function, we asked the question “If you were to spend the rest of your life with your sexual function the way it is today, how would you feel about this?”, with answer choices of “very satisfied”, “somewhat satisfied”, “neither satisfied nor dissatisfied”, “somewhat dissatisfied” and “very dissatisfied”, from part of a questionnaire from the Global Study of Sexual Attitudes and Behaviors (GSSAB) study. The top two categories for each aspect were collapsed to identify positive answers as being very or somewhat satisfied with the level of sexual function. We analyzed relationships between dissatisfaction with sexual function and other variables, including age, stress urinary incontinence, urgency (1⁄4 once a day), daytime frequency ( 8 times/day), and nocturia (1⁄4 once a night). The chi-square test and logistic regression models were used for statistical analyses. Values of P<0.05 were considered statistically significant. RESULTS: A total of 360 individuals completed the questionnaire (response rate, 70.0%). The mean ( standard deviation) age of respondents was 48.3 13.2 years. Prevalences of stress urinary incontinence, urgency, daytime frequency, and nocturia were 35.4%, 3.1%, 39.6%, and 55.0%, respectively. Overall, the prevalence of dissatisfaction with sexual function was 55.4%. In univariate analysis, age, urgency, and nocturia were associated with dissatisfaction with sexual function. In multivariate analysis, a significant correlation was found between dissatisfaction with sexual function and both age (odds ratio (OR), 1.05; p<0.001) and urgency (OR, 9.19; p1⁄40.047). CONCLUSIONS: Our study confirmed age and urgency as independent risk factors for dissatisfaction with sexual function. These results suggest that urgency can offer a predictor of sexual dysfunction among Japanese women.

Chemoprevention and Novel Treatments of Non-Muscle Invasive Bladder Cancer

The Cancer Journal for Clinicians reports there will be 69,250 newly diagnosed cases of bladder cancer in 2011, with 52,020 being men and 17,230 being women with an increase by 50% of annual cases since 1985. Approximately 1 in 5 of those who develop bladder cancer will die due to the disease (relative mortality 20.8%, [Siegel et al., 2011, Golijanin et al., 2006]). Bladder cancer has become the second most prevalent cancer after cancer of the prostate in middle-aged to elderly male individuals. Many patients do not die from their disease, but typically have multiple recurrences (Pelucchi et al., 2006). This lends to a fiveyear cost to Medicare attributed to bladder cancer of over one billion dollars (Yabroff et al., 2008). Tobacco use and exposure to aromatic amines are well established etiologic contributors to bladder cancer and by eliminating or reducing contact with these substances has been shown to reduce such risk. BCG (bacillus Calmette-Guerin) has become the standard of care in the treatment of carcinoma in situ as well as high grade T1 (invasion into the lamina propria) and when not appropriate, Mitomycin-C, has been proven to be an acceptable, albeit, less effective alternate. The goal of this chapter will be to describe novel agents that may show promise in the treatment of bladder cancer. This will include descriptions of the agents, their respective mechanism of action (e.g. molecular/biochemical pathways, cell cycle interaction, necrosis), clinical data, combinations of combinations of regimens and mode of delivery. and mode of delivery. A second goal of this chapter will be to consider whether any of these novel agents may have a role in the prevention of bladder cancer.

MAPK and PI3K kinase inhibition reduces proliferation of Barrett’s adenocarcinoma in vitro

Objectives. Esophageal adenocarcinoma usually arises from Barrett’s esophagus. Mitogen-activated protein kinase (MAPK) and phosphatidylinositol 3-kinase (PI3K) play a critical role in cell survival. We hypothesized that their inhibition in Barrett’s cancer cells would decrease cellular proliferation and alter apoptosis in vitro.Methods. Two Barrett’s-associated adenocarcinoma cell lines, SEG-1 (wild type p53) and BIC-1 (mutant p53), were treated with MAPK (U0126) and PI3K (LY294002) inhibitors at 20 μM concentrations. After 24 and 72 h, cell viability was measured by MTT assay. Apoptosis and necrosis was evaluated by the Annexin V FITC assay. Statistical analysis was performed by ANOVA. Results. LY 294002 and U0126 treatment produced significant reductions (range 15.7 to 62.0%, P < 0.001) in cellular proliferation at both 24 and 72 h in the SEG-1 cells. BIC-1 cell viability was reduced (39.3 to 56.4%, P < 0.001) at 72 h. Both early and late apoptotic activity was significantly increased (P < 0.05) in the SEG-1 cells. Necrosis was significantly reduced (P < 0.01) using both inhibitors. No changes in either early or late apoptosis or necrosis were observed in the BIC-1 cells. Conclusions. Herein, we report the first demonstration of significant anti-proliferative effects against Barrett’s adenocarcinoma by MAPK and PI3 kinase inhibition in vitro. Pro-apoptotic mechanisms prevail in the wild-type p53 cells. Further investigation is warranted to advance the clinical treatment of this devastating disease.

Using alpha-interferon as an adjunctive therapy to reduce the dose of energy required with photodynamic therapy in treating human bladder tumors : Cell culture results

Carcinoma in situ (CIS) of the bladder has long been known as an aggressive killer. Because of the high degree of progression to muscle invasion and the high rate of occult early metastasis, early radical cystectomy had been the preferred treatment, a highly morbid procedure. Even now, with routine use of continent diversions or oithotopic bladder replacements, most patients would agree that there is still a great deal of morbidity associated with this treatment.