bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract A22: Spirometric surveillance for premalignant and malignant bronchial lesions
Authors: Jayaprakash Vijayvel , Loewen Gregory , Dhillon Samjot , Mahoney Martin , Delgado Machare , Menezes Ravi , Bogner Paul , Yendamuri Saikrishna , Hogarth Kyle , Mary Reid
Published: May 2012

Background: Chronic obstructive pulmonary disease (COPD) and lung cancer (LC) are diseases that share common risk factors. It has been reported that more than half of the patients diagnosed with LC also suffer from COPD. Although COPD is a well known risk factor for LC, the relationship between impaired lung function (LF) and the incidence and progression of pre-malignant lesions (PMLs) in the central airways is still unclear.

Methods: The study included 217 high-risk patients from a hospital-based lung cancer surveillance cohort who underwent bronchoscopy with endobronchial biopsy of suspicious lesions, at the Roswell Park Cancer Institute, Buffalo, New York. All patients had lung function measurement within 6 months preceding their baseline biopsy. Baseline histopathology diagnoses included 91squamous metaplasia (SM), 25 squamous dysplasia (SD), 1 in-situ carcinoma and 5 invasive bronchial carcinoma. Follow-up bronchoscopy and biopsy were performed on 69 patients. Sixteen patients had a progression of the baseline lesion to a higher grade. The relationship between the baseline LF measures and the incidence and progression of PMLs were examined using regression models.

Results: Patients with forced expiratory volume in 1 second percent predicted (FEV1%) of <50% had 4.5 times greater risk of being diagnosed with PML/cancer, when compared to patients with FEV1% ≥80. Increased risk was noted for all grades of PMLs and cancers, with the risk being almost 8 times for squamous dysplasia [Odds ratio = 7.95; 95%confidence interval (CI) =1.77–35.6]. Similarly, FEV-1/FVC ratio of <50% was associated with 3 times greater risk of detecting a PML/cancer. When COPD was classified based on GOLD criteria, the patients with severe COPD had 2.7 times and 4.8 times greater risk of being detected with SM and SD, respectively. The risk persisted for SD even when an age standardized classification was used to define COPD. COPD patients also had a non-statistically significant 2.5 times greater risk of their baseline lesion progressing to a higher grade [Hazard Ratio 2.48 (95% CI 0.65 – 9.41), p-value - 0.18)]. The mean time to progression from a lower grade lesion to a higher grade was 27 months for patients with COPD and 50 months for patients without COPD.

Conclusion: Impaired LF can be a good predictor of occurrence and progression of PMLs in central airways of high risk patients. Spirometric measurement of lung function can be used as an additional tool for identifying target populations in need of more aggressive LC surveillance.

Citation Information: Cancer Prev Res 2010;3(1 Suppl):A22.
Abstract A23: PSA testing and prostate cancer mortality in an Asian population: Linking prevalence of elevated PSA and mortality rate
Authors: Chiang Po-Huang , Wen Chi-Pang
Published: May 2012

Background: In Asian populations, prostate cancer mortality (PCM) has been increasing with the increasing use of PSA testing.

Objectives: 1) To establish prevalence of elevated PSA at different cutpoints in Taiwan and to assess the predictive relationship of PSA with PCM. 2) To compare PCM between those with PSA tested and not tested from a cohort study.

Methods: Among the cohort, 231,937, who received standard medical screening examination between 1994 and 2006 at a private screening clinic, 46,951 subjects, aged at least 50 years old, had PSA test results. Age and education were adjusted to those of Taiwan to arrive at national PSA prevalence. PCM was compared between the PSA untested (38,596) and PSA tested group (24,176), with the latter including those with a self-reported history of prostate cancer (1,675).

Results: National prevalence of PSA was 8.8% (threshold at 4) and 1.9% (threshold at 10). With national mortality rate at 30.1/100,000, the chance of dying in ten years was 3.4%, 15.8% or 27.9% among those above threshold at 4, 10, or 15, respectively, yielding false positives rates, with mortality as the outcome, 96.4%, 84.2% or 72.1%, and false negative rates, 12%, 33% or 50%, respectively. Having PSA tested, when compared with not tested, was not associated with decreased mortality from prostate cancer, even though PSA tested group, a higher educational class, had significantly decreased mortality from all cause. Increasing PSA levels were associated with increasing HRs.

Conclusion: PSA value above standard threshold of 4 is common in this Asian population (one out of 11 for age 50 and older), but only one in 29 tested positive was expected to die from prostate cancer within the next 10 years. This 10-year probability increased when threshold was 10 (1/6) but was still low. The high false positive rate of PSA (97%) and the harm associated with intervention among Asians should be communicated to the elderly patients, in view of the particularly low mortality rate of this cancer.

Citation Information: Cancer Prev Res 2010;3(1 Suppl):A23.
Abstract A28: Improving detection through SELF (selective excitation light fluorescence) imaging
Authors: MacAulay Calum , Khojasteh Mehrnoush
Published: May 2012

The use of fluorescence imaging and spectroscopy for the detection of early neoplastic epithelial lesions has been well established, with clinically adopted devices in for use in the lung (Onco-LIFE, SAFE-1000, etc), the oral cavity (VELScope and Identaf 3000) and others in development for the cervix and dermatology. Primarily these imaging technologies make use of fluorescence excitation illumination of a single wavelength (predominately in the near UV to blue wavelength ranges) and differentiate between normal and not normal tissue based upon intensity changes and spectral shifts in the emitted tissue autofluorescence (not normal -darker with less green or blue relative the red fluorescence).

SELF imaging makes use of a multitude of illumination wavelengths to specifically couple to the action spectra of the fluorophores within the tissue to enhance the differentiation between tissue states, fluorophores and their immediate environment. This methodology makes use of the different absorption spectra (action spectra) of different fluorophores or similar flourophores in different environments. Conventionally this can be done through the sequential illumination with many different excitation wavelengths and sequential image capture, to collect a hyperspectral excitation image data cube followed by some form of spectra unmixing to resolve the individual targets (components) contributing to the image. Each target is identified by a unique weighted sum of pixel intensities across the excitation wavelengths in the data cube.

Through the use of a programmable light source such as the OneLight (OneLight Corp.) in which not only the wavelengths of the illumination light, but their individual intensities (alone or in combination) can be selected under computer control it is possible to not only rapidly illuminate with separate excitation wavelengths but to illuminate with a collection of weighted (each wavelength has a different selected intensity) spectra to specifically couple to selected fluorescence targets (specific fluorophores or fluorophores in specific local environments). Thus instead of needing to illuminate with a series of 10 separate excitation wavelengths and collect separate 10 images on can illuminate with a few (2–3) weighted profiles of excitation wavelengths and collect a few (2–3) images, the number of spectra used (images collected) determines the number of targets differentiated. In this fashion it is possible to detect in an image many more specific flourophore types than with conventional fluorescence imaging. SELF imaging in microscopy, wide field macroscopic imaging and ex vivo and in vivo imaging has been demonstrated and will be presented.

Citation Information: Cancer Prev Res 2010;3(1 Suppl):A28.
Abstract A29: Rectal mucosal polyamine and PGE2 levels and risk of colorectal adenomas in a phase IIb/III trial of combination difluoromethylornithine (DFMO) plus sulindac
Authors: Thompson Patricia A. , Wertheim Betsy C. , Zell Jason A. , LeFleur Bonnie , Chen Wen-Pin , McLaren Christine E. , Meyskens Frank L. , Gerner Eugene W.
Published: May 2012

Background: The pharmacologic targeting of polyamines and prostaglandin E2 (PGE2) has proven efficacy for colorectal adenoma prevention. However, the contribution of these analytes as biomarkers of drug response has not been investigated.

Methods: We analyzed the relationships between rectal mucosal levels of polyamines (spermidine, spermine, and putrescine) and PGE2, treatment, and risk of adenoma within the context of a phase IIb/III trial of combination difluoromethylornithine (DFMO) plus sulindac for the prevention of colorectal adenoma.

Results: We detected no differences in the change in PGE2 levels between the two groups for any time points, but we show a statistically significant decrease in spermidine:spermine levels and in putrescine levels in the DFMO/sulindac group at 12 and 36 months compared to the placebo arm. When we asked whether or not change in polyamine or putrescine levels was associated with adenoma recurrence in the treatment arm only, we found no difference in the rate of metachronous events by change in polyamine or PGE2 levels in the treatment arm. We also found no evidence that polyamine or PGE2 levels were associated with risk of adenoma when we restricted the analysis to the placebo arm only. Of interest, we found that participants with low baseline spermidine:spermine levels who received DFMO/sulindac achieved a significantly greater benefit from treatment (RR = 0.15, 95% CI = 0.06 to 0.40) than those with high baseline spermidine:spermine receiving drug (RR = 0.50, 95% CI = 0.27 to 0.92).

Conclusions: DFMO/sulindac significantly suppressed the production of rectal mucosal polyamines, but the magnitude of change was not predictive of response to intervention. This was likely due to the large effect of DFMO/sulindac on adenoma recurrence. We found no evidence in the placebo arm that baseline polyamine or PGE2 were associated with the risk of developing adenoma. The modulating effect of baseline spermidine:spermine levels on metachronous adenoma, in those individuals receiving the drug combination, provides evidence of differential agent-effectiveness by steady-state polyamine levels that should be investigated in future studies.

Citation Information: Cancer Prev Res 2010;3(1 Suppl):A29.
Abstract A3: MicroRNA-related polymorphisms are associated with clinical outcomes in early stage non-small cell lung cancer patients
Authors: Pu Xia , Lu Charles , Stewart David J. , Gu Jian , Hildebrandt Michelle A.T. , Lin Jie , Lippman Scott M. , Wu Xifeng
Published: May 2012

MicroRNAs (miRNA) can function as oncogenes or tumor suppressors. In this study, to test the hypothesis that genetic variation within miRNA processing genes or miRNA binding sites on cancer-related genes may alter clinical outcomes in non-small cell cancer (NSCLC) patients, we genotyped 77 single nucleotide polymorphisms (SNPs) from eight miRNA processing genes and 163 SNPs from 133 predicted miRNA binding sites in 598 patients with early stage (stage I, IIA and IIB) NSCLC to determine the effect of these variations on progression risk and overall survival. We further conducted a subset analysis to exclude potential bias due to different treatment regimens. We identified 37 SNPs associated with overall survival. In the surgery-only patients, 22 SNPs modulated overall survival, including FZD4:rs713065 that remained significant for decreased risk after multiple comparisons (HR: 0.53, 95% CI: 0.36–0.77). For progression risk, 21 and 25 SNPs were found to be significant in the entire and surgery-only patient populations, respectively. Following multiple comparison, ICAM1:rs281437 (HR: 1.80, 95% CI: 1.29–2.52) and MDM4:rs10900596 (HR: 2.01, 95% CI: 1.26–3.19) remained significant increase in progression risk, while FAS:rs2234978 (HR: 0.58, 95% CI: 0.41–0.80) displayed a significantly protective effect. CDC7:rs12125947 (HR: 2.19, 95% CI: 1.37–3.51) remained significant risk effect in the surgery-only patients. A strong cumulative, dose-effect of these variant genotypes to increase risk and dramatically decrease median event-free time was consistently observed. We also identified potential higher-order gene-gene interactions among these SNPs. With validation, our result can be used in the prognosis of clinical outcomes for early stage NSCLC patients.

Citation Information: Cancer Prev Res 2010;3(1 Suppl):A3.
Abstract A30: Cost-effectiveness analysis of HER2 testing methods in adjuvant treatment of HER2 breast cancer
Authors: Ashok Mahima , Griffin Paul , Halpern Michael
Published: May 2012

Background: Trastuzumab is a targeted therapy indicated for the treatment of HER2-postive breast cancers, an aggressive form of this disease. HER2 status can be assessed using two tests: florescence in situ hybridization (FISH), which tests for HER2 gene amplification; or immunohistochemistry (IHC), which tests for HER2 protein overexpression. There is considerable debate regarding which of these tests should be used for HER2 detection due to differences in cost, accuracy, and health outcomes.

Objective: To compare the outcomes and cost effectiveness of FISH and IHC for HER2 testing in breast cancer.

Design: Based on data from outpatient electronic medical records, decision models were designed for FISH, IHC and No Test scenarios (in which all patients received trastuzumab without being tested). Each model considered costs and health outcomes. Treatment possibilities included chemotherapy (with or without trastuzumab), radiation therapy, and surgery. Cost for trastuzumab adverse events were also included. Point estimate analysis and Monte Carlo simulations (varying false positive rates of tests) were used to determine results.

Data: A hypothetical cohort of 100,000 female breast cancer patients. Interventions: Testing with FISH or IHC followed by subsequent trastuzumab treatment when indicated (i.e., when the test result is positive for HER2), which included false positive and false negative results.

Outcome Measures: Medical costs, Quality Adjusted Life Years (QALYs) and percentage of patients correctly tested (true positives who test as positive and true negatives who test as negative).

Results: The expected per-patient costs of the No-test model, IHC model and FISH model were $166,360, $72,405, and $64,626 respectively. The expected values of total QALYs experienced by IHC and FISH groups were essentially identical. Using Monte Carlo analysis to vary test accuracy rates also indicated decreased costs and improved outcomes for FISH. In addition, FISH had increased levels of correct HER2 status detection when compared with IHC: if tested with FISH rather than IHC, there was a 3.58% increase in probability that a patient who is HER2 positive will be diagnosed as positive and a 15.66% increase in probability that a patient who is HER2 negative will be diagnosed as negative.

Conclusions: FISH should replace IHC as the preferred method of choice for HER2 testing. This would result in increased cost-effectiveness of testing and treatment for HER2 breast cancer.

Citation Information: Cancer Prev Res 2010;3(1 Suppl):A30.
Abstract A31: The role of microRNA 125a-3P in the pathogenesis of lung cancer
Authors: Hazra Saswati , Gardner Brian K. , Krysan Kostyantyn , Walser Tonya C. , Brothers John , Larsen Jill E. , Pertsemlidis Alexander , Shay Jerry W. , Minna John D. , Spira Avrum , Dubinett Steven M.
Published: May 2012

Tobacco exposure results in specific oncogene and tumor suppressor gene alterations in lung epithelial cells that often lead to lung cancer development. Activating point mutation of the K-ras proto-oncogene is one of the most frequently occurring genetic changes found in lung cancer both in active and former smokers. Understanding how K-ras mutations mediate the malignant phenotype may yield more effective prevention and diagnosis strategies to be applied to individuals at elevated risk for lung cancer. Micro RNAs are a class of short (19–24 nucleotides) noncoding RNAs that play a critical role in posttranscriptional gene regulation. A rapidly developing literature supports an important role for micro RNAs in the regulation of lung carcinogenesis and tumor progression. In order to better understand the role of micro RNAs in the K-ras-mutated bronchial epithelium, we utilized hTERT- and Cdk4-immortalized human bronchial epithelial cells (HBEC) expressing mutant KrasV12. We next generated the micro RNA profile (TaqMan miRNA array v.2, Applied Biosystems, Foster City, CA), gene expression profile (Affymetrix U133 plus 2 array), and protein expression profile (50-plex inflammatory marker panel) of the K-ras mutated and vector control HBECs. We found that the micro RNA miR-125a-3p was significantly suppressed in K-ras-mutated HBECs compared to the vector control cells. We then evaluated basal expression of miR-125a-3p in both HBECs and NSCLC cell lines containing mutated K-ras by real time RT-PCR. Both K-ras-mutated HBECs and NSCLC cell lines expressed low levels of miR125a-3P. In order to further evaluate the role of miR-125a-3p, we stably expressed this micro RNA in K-ras-mutated HBECs. Our results indicate that miR-125a-3p expression down-regulates several tumor-promoting factors, including Gro-α, HGF, VEGF, GM-CSF, and G-CSF. Further studies revealed that suppression of Gro-α and HGF, but not VEGF, is regulated at the level of mRNA expression. Overexpression of miR-125a-3p suppressed proliferation of the K-ras-mutated HBECs but did not alter the proliferation of the vector control HBECs. Based on these findings, we anticipate that loss of miR-125a-3p may have a broad impact in the bronchial epithelium that harbors K-ras mutations, potentially regulating cellular proliferation, angiogenesis and inflammation. Further exploration is required to evaluate the role of this micro RNA and its downstream effects as potential screening tools and targets of prevention and therapy in the setting of K-ras-mutated lung cancers.

Citation Information: Cancer Prev Res 2010;3(1 Suppl):A31.
Abstract A37: Differential response of LH and hCG on ovarian granulosa cells
Authors: Gupta Chanchal , Chhabra Yash , Chapekar Trimbak , Sinha Subrata , Luthra Kalpana
Published: May 2012

LH hyperstimulation has been linked to conditions like PCOD and granulosa cell tumor. Chapekar had established a model of ovarian tumorigenesis by bilateral ovariectomy and splenic transplantation of ovaries, resulting in sustained LH levels because of absence of feedback inhibition. An in vitro model of the same phenomenon has resulted in goat ovarian granulosa cell line after exposure of primary granulosa cells to LH.

In this study, we have exposed primary ovarian granulosa cells to either hCG or LH in a sustained manner (4 weeks). We observed a marked difference in the morphology and growth pattern between the LH treated, hCG treated and untreated cells. LH treated cells were transparent, spindle shaped with elongated cell extensions. hCG treated cells were distinctly polygonal with shorter cell extensions, while untreated cells were smaller with fewer extensions and lesser in number as compared to the hormone treated cells. Functional characterization of the granulosa cells was done by estimation of progesterone in supernatant by ELISA. Levels of progesterone were similar in all three subsets in primary culture. The untreated and hCG exposed cells showed markedly reduced cell proliferation after the fourth week of exposure while LH treated cells were proliferating rapidly even after 4 weeks. There were marked differences in downstream signaling molecule cAMP after short term (5 days) and long term stimulation (two weeks) with LH and hCG. Intacellular cAMP was assessed by ELISA. On prolonged exposure to LH and hCG, level of cAMP was significantly higher in hCG treated cultures as compared to LH treated cultures. Elevated cAMP levels on hCG stimulation is known to promote apoptosis in cells.

This in vitro model system is suitable to study the effect of sustained hormonal stimulation on granulosa cells and can help in understanding the role of LH and hCG in etiology and pathogenesis of granulosa cell tumor which constitutes 1–2% of ovarian tumors.

Citation Information: Cancer Prev Res 2010;3(1 Suppl):A37.
Abstract A38: Is the mycotoxin zearalenone bioactivated to genotoxic metabolites? In vitro evidence for mercapturic acid formation from catechol metabolites
Authors: Hildebrand Andreas , Pfeiffer Erika , Metzler Manfred
Published: May 2012

The Fusarium toxin zearalenone (ZEN) is a frequent contaminant of corn and other grains world-wide and known for its high estrogenic activity. In a NTP carcinogenicity study, ZEN was found to increase the incidence of hepatocellular adenomas in female mice and of pituitary adenomas in both female and male mice. The mechanism of ZEN tumorigenicity is unknown to date and may be related to the hormonal activity. However, the recent disclosure of a novel pathway in the metabolism of ZEN opens up another possibility: ZEN was found to readily undergo aromatic hydroxylation when incubated with hepatic microsomes from humans and various other mammalian species, and two catechol metabolites were unequivocally identified in our laboratory (Pfeiffer et al., Mol. Nutr. Food Res. 53, 1123–1133, 2009). Thus, the phase I metabolism of ZEN resembles that of the steroidal estrogens 17β-estradiol (E2) and estrone (E1), which also form catechol metabolites. There is increasing evidence that these catechol metabolites contribute to the well-established carcinogenicity of E2 and E1 by acting as genotoxins after further activation to quinones (Cavalieri and Rogan, Ann. N. Y. Acad. Sci. 1089, 286–301, 2006; Bolton and Thatcher, Chem. Rev. Toxicol. 21, 93–101, 2008). A similar mechanism is conceivable for the carcinogenicity of ZEN. In order to obtain evidence for the formation of reactive quinone metabolites from ZEN, it was the aim of the present study to trap such quinones as mercapturic acids in microsomal incubations fortified with N-acetylcysteine (NAC).

Incubations (0.4 mL of 0.1 M phosphate buffer pH 7.4) containing human hepatic microsomes (0.4 mg protein), ZEN (0.1 mM) and a NADPH-generating system were conducted for 10 min at 37°C for the formation of catechol metabolites. 0.1 mL of a 40 mM NAC solution in water was then added and incubation continued for another 30 min. Thereafter, 0.5 mL of 0.7 M glycine/HCl buffer pH 1.2 was added and the NAC adducts were extracted with 2 × 0.5 mL ethyl acetate. Analysis of the extract by LC-MS/MS revealed the presence of at least two NAC adducts, which were identified as the N-acetylcysteinyl derivatives of one of the catechol metabolites of ZEN and of its microsomal reductive metabolite alpha-zearalenol by comparison with authentic synthetic standards.

This is the first report to show that reactive quinone metabolites of ZEN are formed in vitro. The toxicological sequelae of this pathway and its in vivo relevance should now be studied.

Citation Information: Cancer Prev Res 2010;3(1 Suppl):A38.
Abstract A47: The anti-inflammatory role of statins and lung cancer chemoprevention
Authors: Young Robert , Hopkins Raewyn , Eaton Tam
Published: May 2012

Recently published observational studies involving over 450,000 subjects have shown that statins confer protection from lung cancer with a risk reduction of up to 50% compared to those not taking statins (RR=0.50). Statins are known to inhibit the prenylation of GTPases (Rho, Rac, Ras), intracellular signaling molecules involved in upregulation of genes involved in inflammation, lung matrix remodeling and carcinogenic pathways. This pharmacological effect has been shown to occur at normal therapeutic doses. Lung cancer is thought to result from chronic cigarette exposure but occurs in only 10% of smokers. Chronic obstructive pulmonary disease (COPD), characterised by airflow limitation on lung function testing, is found in approximately 60–90% of lung cancer cases and is the most important risk factor for lung cancer in smokers. COPD results from smoke-induced matrix remodeling involving parenchymal destruction (emphysema), airway fibrosis (small airway narrowing) and epithelial proliferation. These processes are believed to be linked to lung cancer through epithelial-mesenchymal transition (EMT), a process whereby matrix remodeling leads to excessive growth factor mediated stimulation of epithelium and subsequent malignant transformation. These premalignant events are mediated by inflammatory cytokines (notably IL-6 and IL-8) released by bronchial epithelial cells following smoke exposure and whose matrix remodeling effects are mediated by GTPases. Statins are known to inhibit the actions of IL-6 and IL-8 by GTPase inhibition thereby inhibiting the upregulating effects of these cytokines on inflammation. Pre-clinical studies show that inhibition of the GTPases can reduce or even reverse EMT. Studies also show that IL-6 is elevated in lung cancer and that anti-IL-6 activity can prevent carcinogensis. Together these observations suggest that statin therapy could, through down regulation of GTPases, inhibit the initiation of lung cancer through inhibition of the upregulatory effects of inflammatory cytokines such as IL-6. Such a mechanism would explain the possible chemo preventive actions of statins, on the development of lung cancer. Further studies are needed to establish the role of statins in inhibiting EMT and malignant transformation leading to lung cancer.

Citation Information: Cancer Prev Res 2010;3(1 Suppl):A47.
Abstract A49: Evidence for gene demethylation in human colon cancer by black raspberries
Authors: Seguin Claire M. , Stoner Kristen , Huang Yi-Wen , Huang Tim , Yang Guang-Yu , Arnold Mark , Martin Edward , Sardo Christine , Stoner Gary D. , Wang Li-Shu
Published: May 2012

Black raspberries (BRBs) exhibit cancer preventative effects in the human colon, in part, by reducing the growth rate and increasing the apoptosis of human colon tumor cells. Recently, we have initiated investigations to determine whether BRBs also cause re-expression of genes in human colon cells that are silenced by aberrant hypermethylation. Cultured human colon cancer cell lines HCT116, Caco2 and SW480, were treated with an anthocyanin-rich (AC) fraction of BRBs every day for five days. AC treatment caused a decreased activity of DNA methyltransferases in a substrate-antibody assay (p<0.05) and a reduction of DNMT1 protein expression. Direct analysis of methylation changes in the promoter region of the suppressor gene p16 (CDKN2A) by Sequenom MassARRAY/EpiTYPER demonstrated a decrease in methylation of all eight p16 promoter CpG units, each unit containing 1–3 CpG sites. All cell lines treated with AC had a significant increase in apoptosis and a decrease in cell proliferation as compared to untreated cells (p< 0.05).

These in vitro observations were confirmed by the examination of normal and tumorous tissue samples taken from 20 colon cancer patients who received daily dietary intervention of freeze-dried BRB powder (60g/day) in drinking water for an average of 3 weeks. DNA from the tissue samples was analyzed by MassARRAY which revealed an overall decrease in p16 methylation in both normal and tumor tissues (p<0.05).

Immunohistochemical analysis of p16 and DNMT1 in the same tissue samples showed that BRB treatment increased p16 expression and decreased DNMT expression (p<0.05), both correlating with decreased methylation of the promoter region of the gene. In addition, the expression levels of Ki-67 and c-myc, both markers of cell proliferation, were reduced in berry treated samples, however, only the reduction in Ki-67 was significant (p<0.05). These data suggest that BRBs are capable of reversing aberrant methylation in both normal and tumorous human colon tissues at tolerable amounts in the diet. This effect appears to be caused by inhibition of DNMT family enzymes, and most likely DNMT1. Supported by NCI grant CA103180 and USDA grant 38903-03560.

Citation Information: Cancer Prev Res 2010;3(1 Suppl):A49.
Abstract A51: Comparison of different berry types to prevent chemically induced papilloma development in the rat esophagus
Authors: Chiu Steve , Seguin Claire , Stoner Kristen , Rocha Claudio , Wang Li-Shu , Stoner Gary D.
Published: May 2012

The present study compared the ability of seven different berry types [black and red raspberries, strawberries, blueberries, acai, noni, goji (wolfberry)] to inhibit N-nitrosomethylbenzylamine (NMBA)-induced tumor development in the F344 rat esophagus. These seven berry types have been shown to differ in their relative content of known chemopreventive agents such as anthocyanins, ellagitannins and carotenoids. Each berry type was picked when ripe, freeze-dried under anoxic conditions to prevent decomposition of their chemical components, and ground into a powder. Beginning at 4 weeks of age, 120 F344 rats were injected with NMBA (0.3 mg/kg, 3x/wk for 5 weeks). Control rats (15) were injected with DMSO/water (80:20), the solvent for NMBA. Beginning one week after treatment with carcinogen, individual berry types were administered at 5% of the diet to groups of 15 NMBA-treated rats until the end of the bioassay (35 weeks). At necropsy, the number of esophageal papillomas was enumerated in all groups of rats. No papillomas were seen in control rats treated with DMSO/water. All berry types caused a significant reduction in the number of NMBA-induced papillomas when compared to rats treated with NMBA only. There was no significant difference in the relative ability of the different berry types to reduce tumor incidence, multiplicity or size. Histopathological studies showed that each berry type was about equally effective in preventing the conversion of preneoplastic lesions (dysplasias) to papillomas. These results suggest that the chemopreventive potential of berries for the rat esophagus is not restricted to black raspberries and strawberries as shown previously in our laboratory. They also suggest that berries that vary markedly in their content of anthocyanins, ellagitannins and carotenoids all exhibit chemopreventive potential in the rat esophagus. Supported by NCI grant CA103180 and USDA grant 38903-03560.

Citation Information: Cancer Prev Res 2010;3(1 Suppl):A51.
Abstract A55: Targeting of FXR and NHE-1 in prevention of esophageal adenocarcinoma
Authors: Zhang Guihong , Ye Fei , Guan Baoxiang , Xu Xiaochun
Published: May 2012

The incidence of esophageal adenocarcinoma, as one of a few human cancers, has been increasing recently in the United States. Tobacco smoke and low fruit and vegetable consumption are risk factors for both esophageal squamous cell carcinoma and adenocarcinoma, whereas frequent gastroesophageal reflux, overweight, and obesity are linked to the increased incidence of esophageal adenocarcinoma. Our research objective is to understand the molecular mechanisms of esophageal carcinogenesis for developing strategies in its early detection and prevention. In our previous studies, we have associated some of these risk factors with altered gene expression in esophageal cancer. We found that benzo[a]pyrene diol epoxide (BPDE, a carcinogen present in tobacco smoke and environmental pollution) and bile acid (a tumor promoter in gastrointestinal cancer) suppressed expression of RAR-β2 and RRIG1, but induced EGFR, p-ERK, AP-1, and COX-2 expression in esophageal cancer cells. In this study, we have detected expression of FXR and NHE-1, a regulator of intracellular pH, in esophageal cancer tissue specimens and then determined effects of their inhibitors in suppressing growth of esophageal cancer cells. We found that expression of FXR and NHE-1 is increased in esophageal cancer compared to the normal tissues. We also found that inhibition of FXR and NHE-1 activity by using guggulsterone and amiloride can effectively suppress the tumor cell growth and gene expression. Further study will reveal the underlying molecular mechanisms and use FXR and NHE-1 expression as biomarkers for early detection or prognosis of esophageal cancer.

Citation Information: Cancer Prev Res 2010;3(1 Suppl):A55.
Abstract A57: High-dose resveratrol modulates drug and carcinogen metabolizing enzymes in a healthy volunteer study
Authors: Chow H-H Sherry , Garland Linda , Hsu Chiu-Hsieh , Vining Donna , Chew Wade , Miller Jessica A. , Perloff Marjorie , Crowell James A. , Alberts David S.
Published: May 2012

Purpose: Resveratrol (RES) or 3, 4′, 5-trihydroxystilbene has been shown to inhibit carcinogenesis by affecting various molecular events in the initiation, promotion and progression stages. The cancer chemopreventive activity of RES has been demonstrated in vivo in a wide variety of tumors including skin, mammary, gastrointestinal, and liver cancer models. Modulation of Phase I and Phase II enzymes has been suggested to be one of the mechanisms responsible for the cancer preventive effect of RES. We conducted a clinical study to determine the effect of pharmacological doses of RES on drug and carcinogen metabolizing enzymes.

Methods: Forty-two healthy volunteers underwent baseline assessment of Phase I and Phase II enzymes. A cocktail of cytochrome P450 (CYP) metabolic probe drugs, including caffeine, dextromethorphan, losartan, and buspirone, were administered to assess the activity of CYP1A2, 2D6, 2C9, and 3A4, respectively. Blood and urine samples were collected for 8 hours after probe drug administration to determine parent probe drug and metabolite concentrations for measurements of CYP enzyme activities. Blood lymphocyte glutathione S-transferase (GST) activity and GST-π level, and serum total and direct bilirubin, a surrogate for UDP-glucuronosyl transferase (UGT) 1A1 activity, were measured to assess Phase II enzymes. After the baseline evaluation, study participants took 1 gm of RES once daily for 4 wks. Enzyme assessment was repeated upon intervention completion.

Results: RES intervention was found to suppress the activity of CYP3A4, 2D6, and 2C9. The geometric mean change of the activity index of CPY3A4, 2D6, and 2C9 was 33% (p = 0.01), 70% (p = 0.01), and 171% (p < 0.0001), respectively. CYP1A2 activity was induced; the geometric mean change of the activity index was 16% (p = 0.005). The overall GST and UGT1A1 activity index were minimally affected by the intervention while an induction of GST-π level (84%, p = 0.002) and UGT1A1 activity index (20%, p = 0.0089) was observed in individuals with baseline enzyme level/activity in the lowest tertile.

Conclusion: We conclude that high doses of RES administration may modulate enzyme systems involved in carcinogen activation and detoxification, which may be one mechanism by which RES inhibits carcinogenesis. However, high doses of RES administration may lead to clinically relevant metabolic drug interactions. Further clinical studies are needed to determine whether lower doses of RES could be used to achieve cancer preventive activities. (Supported by N01CN35158 from the National Cancer Institute, Division of Cancer Prevention)

Citation Information: Cancer Prev Res 2010;3(1 Suppl):A57.
Abstract A6: Ethnic differences in risk of lung cancer associated with repair capacity for tobacco carcinogen-induced DNA adduct
Authors: Wang Li-E , Spitz Margaret R. , Qiao Yawei , Dong Qiong , Amos Christopher I. , Wei Qingyi
Published: May 2012

Lung cancer is initiated by tobacco carcinogens-induced DNA damage, such as DNA adducts caused by benzo[a]pyrene diol epoxide (BPDE). These adducts can be removed by an efficient nucleotide excision repair. We have previously reported that DNA repair capacity (DRC) measured by the host-cell reactivation assay is associated with risk of lung cancer in non-Hispanic whites. In the recent analysis with larger sample sizes including minorities, we found that the suboptimal DRC remains associated with increased risk of lung cancer in 1730 cases and 1683 controls of non-Hispanic whites. The mean values of DRC were 8.37% ± 2.8% (SD) in the cases vs. 9.04% ± 3.1% in the controls (P < 0.0001). The odds ratio (OR) was 1.42 (95% confidence interval [CI], 1.24 – 1.64) after adjustment for age, sex, pack-years of smoking, family history of cancer, and other assay-related covariates. The similar results and risk trend have been also found in 127 Mexican-American patients with lung cancer and 176 cancer-free controls with a mean DRC of 8.38% ± 2.7% in the cases vs. 9.04% ± 3.3% in the controls, and an adjusted OR of 1.67 (95% CI, 1.00 – 2.78), although the difference in mean DRC values was borderline significant (P = 0.056) because of the relatively small sample size. However, this difference in DRC for tobacco carcinogen-induced DNA adducts was not significant in 327 African-American patients with lung cancer and 325 cancer-free controls. The mean values of DRC were 8.90% ± 3.1% in the cases vs. 8.66% ± 2.8% in the controls (P = 0.301). The adjusted OR was 0.99 (95% CI, 0.70 – 1.39). The DRC level in African-American controls was non-significantly lower than other ethnic controls (P = 0.144); however, African-American patients had significantly higher DRC than non-Hispanic white patients and Mexican-Americans patients (P = 0.053). The results further confirmed that low DRC is associated with increased risk of lung cancer in non-Hispanic whites and also provided the evidence of this association in Mexican Americans. These findings help us understand the ethnic differences in the development of lung cancer.

Citation Information: Cancer Prev Res 2010;3(1 Suppl):A6.
Abstract A60: Epigenetic modifications of huntingtin-related genes by dietary components for the prevention and treatment of breast cancer
Authors: Abdolmaleky Sayed Hamid , Eskandari Mohammad , Li Linglin , Zhou Jin-Rong
Published: May 2012

Huntingtin protein (Htt) interplays with a large numbers of proteins and participates in diverse cellular pathways. Wide variety of Htt binding partners have been identified, including the Htt-Interacting protein-1 (HIP1) and Htt-associated protein-1 (HAP1). Htt has been shown pro-apoptotic activity, and malfunctioning of Htt may result in cell outgrowth. HIP1 is involved in endocytosis and receptor trafficking and introduced as an oncoprotein. An altered expression of HIP1 has been reported in various cancers including breast cancer. HAP1 maintains the normal level of membrane TrkA by preventing the degradation of internalized TrkA. Recent evidence supports a role of HAP1 in micro-tubule-dependent organells transport, but its role in cancer development and progression has not been studied. We propose that the HAP1 and HIP1 may serve as a pair of “Ying-Yang” modulators on the Htt functions and that the Htt-HAP1-HIP1 pathway may play an important role in the development and progression of breast cancer. We further propose that dietary/nutritional components that target this pathway may have chemopreventive effects on breast cancer. The objective of this study was to evaluate the differential expression and epigenetic modification patterns of Htt pathway related genes in normal and cancerous breast cells treated by dietary active components.

Human breast cancer cells were treated with dietary components known to prevent cancer cell growth and the effects of these components on HAP1, HIP1 and HTT expressions and promoter DNA methylations were investigated by using real-time PCR, Methylation Specific PCR (MSP) and bisulfite sequencing. HAP1 gene was silenced in several breast cancer cell lines including MCF7, MDA-MB231, MB453, SKBR3 and in much lower extent in immortalized MCF10A cells, but not in normal human mammary epithelial cell line (HMEC). Epigenetic analysis for Htt, HIP1 and HAP1 showed severe HAP1 promoter DNA hypermethylation in breast cancer cell lines, but not in HMEC. When the cells were treated with 5-AZaC (a DNA demethylating agent) and butyrate (a HDAC inhibitor), we found that 5-AZaC treatment significantly increased HAP1 expression by over 100 times, but butyrate had little effects. Treatment of the cells with dietary active compounds, sulfuraphane, EGCG, oridonin, tanshinone 2A and cryptotanshinone increased expression of HAP1 and HTT genes and decreased expression of HIP1 gene in HMEC, more significantly in cancer cell lines MCF7 and MB453. Preliminary studies indicate that these dietary components acted through modulation of HAP1 promoter methylation and further experiments are taking place to construct HTT and HAP-1 expressing plasmids for further in vitro and in vivo gene function studies.

Citation Information: Cancer Prev Res 2010;3(1 Suppl):A60.
Abstract A62: Increased histone acetylation, but no inhibition of HDAC activity, by kale sprout intervention in a human prostate cancer xenograft model
Authors: Kim Jung-Hyun , Bähr Marion , Wagner Julia , De Nicola Gina , Rüfer Corinna , Jung Manfred , Iori Renato , Gerhäuser Clarissa
Published: May 2012

Cruciferous vegetable consumption has been associated with a reduced risk of prostate cancer. These vegetables such as broccoli and kale are rich in glucosinolates, which are converted to isothiocyanates (ITCs) by myrosinase. ITCs have cancer preventive potential through various mechanisms including anti-proliferative effects. ITC metabolites have been recently identified as inhibitors of histone deacetylases (HDAC), which have the potential to turn on epigenetically silenced genes, leading to cell cycle arrest and apoptosis.

In the present study we investigated the effect of kale sprouts commercially available as a food supplement to inhibit the growth of human LNCaP prostate cancer cells in a xenograft model. We were interested whether kale sprout intervention would affect histone acetylation and in studying underlining mechanisms, e.g. HDACs and histone acetyltransferases (HAT). LNCaP human prostate cancer cells were injected to male nude Balb/c mice (10 per group). Mice were fed with either regular rodent chow or chow supplemented with 20% kale sprouts (containing about 60 µmol glucosinolates per 5 g chow consumed daily, together with active myrosinase) from a week before cell injection until sacrifice. 7.5 weeks after injection, tumor tissue was collected.

Kale sprout intervention did not significantly inhibit tumor growth in the xenograft model. However, global acetylation levels of histone H4 detected by Western blotting were increased in the kale sprout-treated group, whereas acetylated histone H3 levels were not significantly changed. The increase of H4 lysine 5, 8, 12 and 16 acetylation was individually confirmed. Among them, acetylated H4 K12 was most abundant. Even though global acetylation of H3 did not change, acetylated H3K9 was significantly increased. Unexpectedly, HDAC activity in nuclear extracts of tumor tissue determined by using ZMAL substrate was not significantly inhibited in tumor tissue by kale sprout intervention. Also, protein expression of HDAC family members HDAC1-5 was not changed overall, although HDAC 5 expression was reduced while HDAC2 expression was increased. Instead, kale sprout treatment significantly increased mRNA levels of PCAF and CBP quantified by qRT-PCR, by 36% and 23%, respectively, in tumor tissue (p<0.001), which may contribute to increased H4 acetylation.

Taken together, our results indicate that dietary consumption of kale sprouts increased global acetylation of histone H4 in LNCaP prostate cancer xenograft model, possibly by induction of HAT expression rather than inhibition of HDAC activity.

Citation Information: Cancer Prev Res 2010;3(1 Suppl):A62.
Abstract A63: Superoxide-dependent MKP-1 degradation and JNK activation contribute to luteolin-induced cytotoxicity in lung cancer cells
Authors: Bai Lang , Xu Shanling , Wang Xia , Ju Wei , Chen Wenjie , Lin Yong
Published: May 2012

Epidemiological studies have suggested luteolin (3′,4′,5,7-tetrahydroxyflavone) as a potential cancer preventive agent. Recently we found that luteolin selectively kills lung cancer but not immortalized human normal bronchial epithelial cells (HBECs). To elucidate the mechanism of luteolin's cancer cell killing activity, we used an in vitro cell culture system to investigate the cytotoxic cellular signaling pathway in lung cancer cells. The results show that luteolin potently induced both apoptosis and necrosis, which was associated with mitochondrion-dependent superoxide accumulation in lung cancer cells. The c-Jun N-terminal kinase (JNK) was substantially activated in luteolin treated cancer cells and luteolin-induced cytotoxicity was effectively blocked when JNK was inhibited. The reactive oxygen species scavengers BHA and NAC, which effectively block luteolin-induced superoxide accumulation, potently suppressed luteolin-induced JNK activation and cytotoxicity, suggesting that luteolin's lung cancer cell killing requires superoxide-mediated JNK activation. Furthermore, JNK activation was tightly associated with a superoxide-dependent rapid reduction of the JNK inactivating phosphatase MKP-1, which was mainly due to destabilizing MKP-1 protein. These results suggest that inhibiting MKP-1 is the main mechanism for luteolin-induced JNK activation. Thus, luteolin activates a novel cytotoxic pathway consisting of superoxide, MKP-1 and JNK in lung cancer cells. Interestingly, although luteolin-induced superoxide accumulation was comparable to that in lung cancer cells, it failed in inducing JNK activation and cell death in the HBECs. Therefore, luteolin-induced cellular signaling in normal and lung cancer cells diverges at the step of JNK activation. Taken together, our results suggest that activation of the speroxide/MKP1/JNK pathway underlies the main mechanism of luteolin's selective ctyotoxicity in lung cancer cells and manipulation of this pathway could be a useful approach for applying luteolin for lung cancer prevention and therapy.

Citation Information: Cancer Prev Res 2010;3(1 Suppl):A63.