bluuebunny/crossref_metadata_2025 huggingface dataset:
4-Aminobiphenyl-Hemoglobin Adducts and Risk of Smoking-Related Disease in Never Smokers and Former Smokers in the European Prospective Investigation into Cancer and Nutrition Prospective Study
Authors: Airoldi Luisa , Vineis Paolo , Colombi Alessandro , Olgiati Luca , Dell'Osta Carlo , Fanelli Roberto , Manzi Luca , Veglia Fabrizio , Autrup Herman , Dunning Alison , Garte Seymour , Hainaut Pierre , Hoek Gerard , Krzyzanowski Michal , Malaveille Christian , Matullo Giuseppe , Overvad Kim , Tjonneland Anne , Clavel-Chapelon Francoise , Linseisen Jakob , Boeing Heiner , Trichopoulou Antonia , Palli Domenico , Peluso Marco , Krogh Vittorio , Tumino Rosario , Panico Salvatore , Bueno-De-Mesquita Hendrik B. , Peeters Petra H. , Lund Eiliv , Agudo Antonio , Martinez Carmen , Dorronsoro Miren , Barricarte Aurelio , Chirlaque M. Dolores , Quiros Josè R. , Berglund Goran , Järvholm Bengt , Hallmans Goran , Day Nicholas E. , Allen Naomi , Saracci Rodolfo , Kaaks Rudolf , Riboli Elio
Published: September 2005

The aim of this study was to evaluate whether biomarkers of environmental tobacco smoke exposure [i.e., 4-aminobiphenyl-hemoglobin (4-ABP-Hb) adducts] were predictive of the risk of tobacco-related cancers and diseases. We did a case-control study nested within the European Prospective Investigation into Cancer and Nutrition, involving 190 controls and 149 cases (incident cancer of the lung, bladder, pharynx, larynx, oral cavity, leukemias, and chronic obstructive pulmonary disease or emphysema deaths). All individuals were never smokers or ex smokers for >10 years. 4-ABP-Hb adducts were analyzed in peripheral blood collected before the onset of the disease (median, 7 years). Overall, 4-ABP-Hb adducts were higher, although not statistically significantly so, in cases (as a whole) than controls. In the control population, high fruit and vegetable consumption significantly lowered the frequency of detectable adducts (Fisher's exact test, P = 0.025). Restricting the analysis to women, 4-ABP-Hb adducts were higher in cases than controls (Mann-Whitney P = 0.036) and the odds ratio (OR) for the presence/absence of adducts was 2.42 [95% confidence interval (95% CI), 1.18-4.98]. Moreover, the association of adducts with the individual cancer types was stronger in women than in the whole study population, although statistically significant only for leukemias (OR, 2.77; 95% CI, 1.06-7.20). The results provide some evidence that women may be more susceptible to environmental tobacco smoke, as suggested by their higher adduct levels. The most important finding of this prospective study is that, at least in women, 4-ABP-Hb adducts may help identify subjects at high risk of cancers related to environmental tobacco smoke exposure.
Evidence for Selective Expression of the p53 Codon 72 Polymorphs: Implications in Cancer Development
Authors: Siddique Mobin M. , Balram Chowbay , Fiszer-Maliszewska Lucja , Aggarwal Amit , Tan Angie , Tan Patrick , Soo Khee C. , Sabapathy Kanaga
Published: September 2005

Polymorphism at codon 72 of p53 results in either the arginine or proline form of p53, whose functional significance in carcinogenesis is controversial. We have investigated if the expression of these p53 polymorphs is selectively regulated, using mRNA from peripheral blood of healthy Asian (Chinese) and the Caucasian (Polish) arginine/proline (arg/pro) heterozygote subjects. Asians were found to preferentially express the pro allele whereas the Caucasians preferentially express the arg allele. On the contrary, about 75% of the heterozygote Chinese breast cancer patients preferentially expressed the arg allele, which rarely contained any somatic mutations. Moreover, histologically normal tissues from Chinese heterozygote breast cancer patients showed selective expression of the arg allele, in contrast to the preferential expression of the pro allele in heterozygote healthy normal breast tissues. Together, the data suggest that the expression of the different p53 polymorphs is selectively regulated in different ethnic populations, and that the arg allele is activated during cancer development in Asians. Thus, the expression status of the p53 polymorphs, rather than the genotypic status, might be a useful indicator for cancer susceptibility.
Selected Genetic Polymorphisms in MGMT, XRCC1, XPD, and XRCC3 and Risk of Head and Neck Cancer: A Pooled Analysis
Authors: Huang Wen-Yi , Olshan Andrew F. , Schwartz Stephen M. , Berndt Sonja I. , Chen Chu , Llaca Victor , Chanock Stephen J. , Fraumeni Joseph F. , Hayes Richard B.
Published: September 2005

Tobacco and alcohol consumption are the major risk factors for head and neck cancer, likely due to DNA-damaging processes. Genetic variations in DNA repair genes may affect an individual's susceptibility to head and neck cancer. Pooling data and DNA specimens from three case-control studies in western Washington State, North Carolina, and Puerto Rico, totaling 555 cases (430 whites) and 792 controls (695 whites), we studied the risk of head and neck cancer in relation to common nonsynonymous single-nucleotide polymorphisms in four DNA repair genes: MGMT (Leu84Phe and Ile143Val), XRCC1 (Arg399Gln), XPD (Lys751Gln), and XRCC3 (Thr241Met). All single-nucleotide polymorphisms were assayed in a single laboratory. Among whites, carriage of the MGMT Phe84 [odds ratio (OR), 0.71; 95% confidence interval (95% CI), 0.51-0.98] or Val143 (OR, 0.66; 95% CI, 0.47-0.92) allele was associated with a decreased risk of head and neck cancer; the haplotype distribution for MGMT differed significantly between cases and controls (covariate-adjusted global permutation test, P = 0.012). The XRCC1 GlnGln399 genotype was also associated with decreased risk among whites (OR, 0.56; 95% CI, 0.32-0.94), whereas XPD751 and XRCC3241 were not associated with risk. Alcohol-related risks tended to vary with DNA repair genotypes, especially for MGMT variants, whereas no effect modification was noted with tobacco use. Consistent findings from three case-control studies suggest that selected DNA repair enzymes may play a role in head and neck carcinogenesis.
Improving Organochlorine Biomarker Models for Cancer Research
Authors: Wolff Mary S. , Britton Julie A. , Teitelbaum Susan L. , Eng Sybil , Deych Elena , Ireland Karen , Liu Zhisong , Neugut Alfred I. , Santella Regina M. , Gammon Marilie D.
Published: September 2005

Multivariate methods were used to predict levels of dichlorodiphenyldichloroethene (DDE) and polychlorinated biphenyl (PCB) concentrations in plasma from characteristics that included age, diet, race, reproductive history, socioeconomic status, and reported body mass index (BMI) at several decades of life before blood collection. Measurements were available for organochlorine compound (organochlorines), cholesterol, and triglycerides in plasma from 1,008 women participants in a population-based case-control study of breast cancer undertaken in 1996 to 1997 on Long Island, NY. Organochlorine compound levels were associated with age, race, lactation history, body size characteristics, and plasma lipids. PCB predictors also included fish consumption. DDE was correlated with current BMI, BMI at every decade of age from ages 20 to 60 years, and BMI-gain (from ages 20 or 30 years to 1997). In contrast, PCBs were correlated inversely with both BMI (fifth to seventh decades of age) and BMI-gain. After adjusting for covariates, DDE and PCB were both positively associated with BMI and inversely with BMI-gain; they were lowest with low BMI, high BMI-gain, and longer lactation. This pattern is consistent with a pharmacokinetic model that predicts higher body burdens during windows of highest uptake, faster elimination of organochlorine compounds in leaner women, and lowered levels accompanying BMI-gain. As a result, lifetime intake for specific organochlorine compound may lead to different plasma levels dependent on changes in body size, absolute intensity of intake, and whether exposure is ongoing (i.e., PCB) or long discontinued (i.e., DDE).
Total and Percent Free Prostate-Specific Antigen Levels among U.S. Men, 2001-2002
Authors: Saraiya Mona , Kottiri Benny J. , Leadbetter Steven , Blackman Don , Thompson Trevor , McKenna Matthew T. , Stallings Fred L.
Published: September 2005

Background: Because total prostate-specific antigen (PSA) and, more recently, the percent free PSA are used to screen men for prostate cancer, population-based, age- and race-specific distributions are needed of both PSA tests among American men to estimate the effect of lowering the PSA threshold or widespread introduction of the free PSA test as an additional screening test.

Methods: We did PSA assays on serum samples from men of ages 40 years and older (n = 1,320) who participated in the 2001-2002 National Health and Nutrition Examination Survey.

Results: About 6.1% (95% confidence interval, 4.7-7.7%), corresponding to an estimated 3.4 million (range, 2.7-4.3 million) men nationwide, ages 40 years and older, had a total PSA of >4.0 ng/mL. Among men ages 50 to 69 years old, the age group for which PSA testing is most prevalent, 5.4% or an estimated 900,000 to 2 million men had a total PSA of >4.0 ng/mL. An equal number had a total PSA between 2.5 and 4.0 ng/mL and a percent free PSA of <25%. Approximately 27% of men in this age group, corresponding to a range of 5.7 to 8.1 million men, had a total PSA <2.5 ng/mL and a percent free PSA of <25%.

Conclusion: The effect of lowering the total PSA threshold or introducing another screening test is significant. Provision of the number of U.S. men with certain total PSA and percent free PSA values may help guide prostate cancer public health policy and screening practices.
Vegetable and Fruit Intake and Pancreatic Cancer in a Population-Based Case-Control Study in the San Francisco Bay Area
Authors: Chan June M. , Wang Furong , Holly Elizabeth A.
Published: September 2005

Pancreatic cancer is one of the most devastating and rapidly fatal cancers, yet little is known about the primary cause and prevention of this disease. We conducted a population-based case-control study to investigate the association between vegetables and fruits and pancreatic cancer. Between 1995 and 1999, 532 cases and 1,701 age- and sex-matched controls completed direct interviews using a semiquantitative food-frequency questionnaire. No proxy interviews were conducted. We observed inverse associations between consumption of total and specific vegetables and fruits and the risk of pancreatic cancer. The odds ratio and 95% confidence interval for the highest versus the lowest quartile of total vegetable intake was 0.45 (0.32-0.62), trend P < 0.0001; and for total fruits and fruit juice was 0.72 (0.54-0.98), trend P = 0.06. Odds ratios and 95% confidence intervals for the highest versus the lowest quartile of specific vegetables and fruits were: 0.63 (0.47-0.83) for dark leafy vegetables, 0.76 (0.56-1.0) for cruciferous vegetables, 0.59 (0.43-0.81) for yellow vegetables, 0.56 (0.41-0.76) for carrots, 0.51 (0.38-0.70) for beans, 0.46 (0.33-0.63) for onions and garlic, and 0.78 (0.58-1.0) for citrus fruits and juice. Compared with less than five servings per day of total vegetables and fruits combined, the risk of pancreatic cancer was 0.49 (0.36-0.68) for more than nine servings per day. These results suggest that increasing vegetable and fruit consumption, already recommended for the prevention of several other chronic diseases, may impart some protection against developing pancreatic cancer.
Incidence Trends of Adenocarcinoma of the Cervix in 13 European Countries
Authors: Bray Freddie , Carstensen Bendix , Møller Henrik , Zappa Marco , Žakelj Maja Primic , Lawrence Gill , Hakama Matti , Weiderpass Elisabete
Published: September 2005

Rapid increases in cervical adenocarcinoma incidence have been observed in Western countries in recent decades. Postulated explanations include an increasing specificity of subtype—the capability to diagnose the disease, an inability of cytologic screening to reduce adenocarcinoma, and heterogeneity in cofactors related to persistent human papillomavirus infection. This study examines the possible contribution of these factors in relation with trends observed in Europe. Age-period-cohort models were fitted to cervical adenocarcinoma incidence trends in women ages <75 in 13 European countries. Age-adjusted adenocarcinoma incidence rates increased throughout Europe, the rate of increase ranging from around 0.5% per annum in Denmark, Sweden, and Switzerland to ≥3% in Finland, Slovakia, and Slovenia. The increases first affected generations born in the early 1930s through the mid-1940s, with risk invariably higher in women born in the mid-1960s relative to those born 20 years earlier. The magnitude of this risk ratio varied considerably from around 7 in Slovenia to almost unity in France. Declines in period-specific risk were observed in United Kingdom, Denmark, and Sweden, primarily among women ages >30. Whereas increasing specificity of subtype with time may be responsible for some of the increases in several countries, the changing distribution and prevalence of persistent infection with high-risk human papillomavirus types, alongside an inability to detect cervical adenocarcinoma within screening programs, would accord with the temporal profile observed in Europe. The homogeneity of trends in adenocarcinoma and squamous cell carcinoma in birth cohort is consistent with the notion that they share a similar etiology irrespective of the differential capability of screen detection. Screening may have had at least some impact in reducing cervical adenocarcinoma incidence in several countries during the 1990s.
p53 Genotypes and Risk of Glioma and Meningioma
Authors: Malmer Beatrice , Feychting Maria , Lönn Stefan , Ahlbom Anders , Henriksson Roger
Published: September 2005

Brain tumors have previously been associated with the Li-Fraumeni syndrome that often is caused by p53 germ line mutations. Therefore, we investigated if polymorphisms of p53 were associated with an increased risk of meningioma and glioma and integrated the polymorphism analyses with detailed information on family history of cancer. In a population-based case-control study, DNA was extracted from 205 glioma and 164 meningioma cases identified during 2000 to 2002 in Sweden and from 374 controls selected randomly from the general population, stratified on age, sex, and geographic region. The Swedish Cancer Registry confirmed a cancer in family members in 86% of cases and controls that reported a family history of cancer. p53 single nucleotide polymorphism (SNP) analyses were done on three SNPs from the promoter region, codon 72 in exon 4, and intron 6. Overall, no associations were found for any of the SNPs. Analyses of the combinations of the three SNPs were also done. The CC-CG-CC-specific polymorphism combination was associated with an odds ratio (OR) of 1.36 [95% confidence interval (95% CI), 0.68-2.72] for glioma and 1.36 (0.64-2.88) for meningioma. When restricting the analyses to cases and controls with a positive family history of cancer, the corresponding results were OR of 3.62 (95% CI, 1.05-12.48) for glioma and 5.69 (1.81-17.96) for meningioma. This study is novel in suggesting an increased risk of brain tumors when the analysis is restricted to those with a history of cancer in the family. However, we cannot rule out the possibility that these results are due to chance.
Insulin Resistance, Apoptosis, and Colorectal Adenoma Risk
Authors: Keku Temitope O. , Lund Pauline Kay , Galanko Joseph , Simmons James G. , Woosley John T. , Sandler Robert S.
Published: September 2005

Compelling evidence from epidemiologic studies indicates that elevated circulating insulin-like growth factor (IGF)-I, insulin resistance, and associated complications, such as elevated fasting plasma insulin, glucose and free fatty acids, glucose intolerance, increased body mass index, and visceral adiposity, are linked with increased risk of colorectal cancer. However, the role of insulin and markers of glucose control in the development of adenomas, precursors to colorectal cancer, has not been fully explored. We evaluated the relationship between plasma insulin, glucose, IGF-I, IGF-II, IGF-binding protein-3 (IGFBP-3), apoptosis, and colorectal adenomas in a case-control study. Participants were drawn from consenting patients undergoing colonoscopy at the University of North Carolina hospitals (Chapel Hill, NC). Participants were classified as cases or controls based on whether they had one or more colorectal adenomatous polyps. Fasting plasma insulin, IGF-I, IGF-II, and IGFBP-3 levels were assessed by ELISA. Glucose was measured by glucose hexokinase assay. Apoptosis was assessed by morphology on H&E-stained sections. Dietary and lifestyle information were obtained by telephone interview. Logistic regression was used to examine the association between adenoma status and insulin-IGF markers. Adenoma cases (n = 239) and adenoma-free controls (n = 517) provided rectal biopsies and/or blood samples and interview data. Consistent with prior findings, cases were more likely to be males, older, have higher waist-to-hip ratio, lower calcium intake, lower apoptosis, and less likely to report nonsteroidal anti-inflammatory drug use. Those in the highest quartile of insulin (adjusted odds ratio, 2.2; 95% confidence interval, 1.1-4.2) and glucose (adjusted odds ratio, 1.8; 95% confidence interval, 0.9-3.6) were more likely to have an adenoma compared with the lowest quartile. Similarly, subjects in the highest two quartiles of insulin were more likely to be in the lowest two quartiles of apoptosis. Overall, there were no significant differences between mean circulating levels of glucose, IGF-I, IGF-II, and IGFBP-3 among cases and controls and no association between these variables and apoptosis. The results provide novel evidence that elevated insulin and glucose are associated with increased adenoma risk and decreased apoptosis in normal rectal mucosa. These findings suggest that insulin may act early in the adenoma-carcinoma sequence to promote the development of colorectal adenoma by decreasing apoptosis in the normal mucosa.
Relative Validity of a Food Frequency Questionnaire with a Meat-Cooking and Heterocyclic Amine Module
Authors: Cantwell Marie , Mittl Beth , Curtin Jane , Carroll Ray , Potischman Nancy , Caporaso Neil , Sinha Rashmi
Published: September 2005

The nutrient and heterocyclic amine (HCA) intake of 165 healthy participants was assessed using a self-administered food frequency questionnaire (FFQ), which included a meat-cooking practices module. A database containing the HCA [2-amino-3,8-dimethylimadazo [4,5-f] quinoxaline (MeIQx) and 2-amino-1-methyl-6-phenylimadazo [4,5-b] pyridine (PhIP)] composition of various types of meat, cooked by different methods and to varying degrees, was developed and validated in conjunction with this module. The relative validity of dietary and HCA intake estimated by the FFQ was investigated using multiple food diaries (3 sets of 4 nonconsecutive day diaries completed over a 3-month period) as the reference method. Crude correlation coefficients of HCA intake assessed by the FFQ and food diaries were 0.43 [95% confidence interval (CI) 0.30–0.55] for MeIQx and 0.22 (95% CI 0.07–0.36) for PhIP intake. Deattenuated correlations were 0.60 (95% CI 0.49–0.69) and 0.36 (95% CI 0.22–0.49), respectively. Absolute MeIQx and PhIP intake was, however, underestimated by the FFQ (21.9 and 78.1 ng/day) compared with the food diaries (34.9 and 263.8 ng/day). The FFQ underestimated total red meat intake, the percentage of consumers, and the median intake of roast/baked and microwaved red meat. PhIP intake was severely underestimated by the FFQ and was most likely because of an underestimation of the percentage of people who cooked chicken using PhIP-producing cooking methods such as broiling and pan-frying. Additionally, the FFQ overestimated the percentage of consumers of baked chicken, a cooking method that produces less PhIP. In conclusion, although the FFQ and meat module underestimated absolute MeIQx and PhIP intake, its ability to rank individuals according to intake was acceptable.
Prognostic Significance of p27kip-1 Expression in Colorectal Adenocarcinomas Is Associated with Tumor Stage
Authors: Manne Upender , Jhala Nirag C. , Jones Jennifer , Weiss Heidi L. , Chatla Chakrapani , Meleth Sreelatha , Suarez-Cuervo Catalina , Grizzle William E.
Published: September 2005

Purpose: Although the decreased expression of p27kip-1, a cyclin-dependent kinase inhibitor, has been correlated with advanced tumor stage and short survival of patients with colorectal adenocarcinomas (CRCs), its prognostic value based on the tumor site, tumor stage, and patient ethnicity was not assessed. Therefore, in this study, we investigated whether the prognostic value of p27kip-1 expression varies with the tumor site, tumor stage and patient ethnicity.

Experimental Design: We evaluated 206 (85 African Americans and 121 Caucasians) archival tissue specimens of first primary CRCs for immunohistochemical expression of p27kip-1, and its prognostic significance was analyzed using univariate Kaplan-Meier and multivariate Cox regression survival methods.

Results: Although, similar proportion of CRCs with decreased p27kip-1 expression was observed in all stages (range, 26–36%), the decreased p27kip-1 expression has been shown as a marker of poor prognosis only for patients with stage III tumors both in univariate (log-rank test, P = 0.014) and multivariate (hazard ratio = 3.2, 95% confidence interval = 1.3–7.7; P = 0.01) survival analyses. The decreased expression of p27kip-1 was associated with a high histologic grade (P = 0.016) in stage II CRCs, and with distal tumors (P = 0.001), tumor invasion (P = 0.044), and with local recurrence (P = 0.008) in stage III CRCs.

Conclusions: No prognostic significance was found for p27kip-1 expression in stages I, II, or IV CRCs, and its prognostic value was not associated with either ethnicity or tumor location. These studies suggest that decreased expression of p27kip-1 is an indicator of poor prognosis and aids in identifying a subset of patients with aggressive forms of stage III CRCs.
High Specificity of Quantitative
Excision Repair Cross-Complementing 1
Messenger RNA Expression for Prediction of Minor Histopathological Response to Neoadjuvant Radiochemotherapy in Esophageal Cancer
Authors: Warnecke-Eberz Ute , Metzger Ralf , Miyazono Futoshi , Baldus Stephan E. , Neiss Susanne , Brabender Jan , Schaefer Hartmut , Doerfler Walter , Bollschweiler Elfriede , Dienes Hans P. , Mueller Rolf P. , Danenberg Peter V. , Hoelscher Arnulf H. , Schneider Paul M.
Published: September 2005

Purpose: The excision repair cross-complementing 1 (ERCC1) gene is coding for a nucleotide excision repair protein involved in the repair of radiation- and chemotherapy-induced DNA damage. We examined the potential of quantitative ERCC1 mRNA expression to predict minor or major histopathological response to neoadjuvant radiochemotherapy (cisplatin, 5-fluorouracil, and 36 Gy of radiation) followed by transthoracic en bloc esophagectomy in patients with locally advanced esophageal cancer (cT2–4, Nx, M0).

Experimental Design: Tissue samples were collected by endoscopic biopsy before treatment. RNA was isolated from biopsies, and quantitative real-time reverse transcriptase PCR assays were performed to determine ERCC1 mRNA expression. Relative mRNA levels (tumor/normal ratios) were calculated as (ERCC1/β-actin in tumor)/(ERCC1/β-actin in paired normal tissue). ERCC1 expression levels were correlated with the objective histopathological response in resected specimens. Histomorphological regression was defined as major response when resected specimens contained <10% of residual vital tumor cells or in case a pathologically complete response was achieved.

Results: Twelve of 36 tumors showed a major histopathological response, and 24 of 36 showed a minor histopathological response. Relative expression levels of ERCC1 of >1.09 were not associated with a major histopathological response (sensitivity, 62.5%; specificity, 100%) and 15 of 24 patients with minor histopathological response to the delivered neoadjuvant radiochemotherapy could be unequivocally identified. This association of dichotomized relative ERCC1 mRNA expression and histopathological response was statistically significant (P < 0.001).

Conclusions: Relative expression levels of ERCC1 mRNA determined by quantitative real-time reverse transcriptase-PCR appear highly specific to predict minor response to our neoadjuvant radiochemotherapy protocol in patients with locally advanced esophageal cancer and could be applied to prevent expensive, noneffective, and potentially harmful therapies in a substantial number (42%) of patients.
Very High Frequency of Hypermethylated Genes in Breast Cancer Metastasis to the Bone, Brain, and Lung
Authors: Mehrotra Jyoti , Vali Mustafa , McVeigh Megan , Kominsky Scott L. , Fackler Mary Jo , Lahti-Domenici Jaana , Polyak Kornelia , Sacchi Nicoletta , Garrett-Mayer Elizabeth , Argani Pedram , Sukumar Saraswati
Published: September 2005

Purpose: Most often it is not the primary tumor, but metastasis to distant organs that results in the death of breast cancer patients. To characterize molecular alterations in breast cancer metastasis, we investigated the frequency of hypermethylation of five genes (Cyclin D2, RAR-β, Twist, RASSF1A, and HIN-1) in metastasis to four common sites: lymph node, bone, brain, and lung.

Experimental Design: Methylation-specific PCR for the five genes was performed on DNA extracted from archival paraffin-embedded specimens of paired primary breast cancer and its lymph nodes (LN) metastasis (n = 25 each); in independent samples of metastasis to the bone (n = 12), brain (n = 8), and lung (n = 10); and in normal bone, brain, and lung (n = 22).

Results: No hypermethylation was detected in the five genes in the normal host tissues. In paired samples, LN metastasis had a trend of higher prevalence of methylation compared with the primary breast carcinoma for all five genes with significance for HIN-1 (P = 0.04). Compared with the primary breast carcinomas, all five genes had higher methylation frequencies in the bone, brain, and lung metastasis, with HIN-1 and RAR-β methylation being significantly higher (P < 0.01) in each group. Loss of expression of all five genes correlated, with a few exceptions, to hypermethylation of their promoter sequences in metastatic carcinoma cells microdissected from LNs.

Conclusion: The frequent presence of hypermethylated genes in locoregional and distant metastasis could render them particularly susceptible to therapy targeted toward gene reactivation combining demethylating agents, histone deacetylase inhibitors, and/or differentiating agents.
Analysis of Aberrant DNA Methylation and Human Papillomavirus DNA in Cervicovaginal Specimens to Detect Invasive Cervical Cancer and Its Precursors
Authors: Widschwendter Andreas , Gattringer Conny , Ivarsson Lennart , Fiegl Heidi , Schneitter Alois , Ramoni Angela , Müller Hannes M. , Wiedemair Annemarie , Jerabek Susanne , Müller-Holzner Elisabeth , Goebel Georg , Marth Christian , Widschwendter Martin
Published: September 2005

Purpose: Cancer of the uterine cervix is an important cause of death in women worldwide. Pap smears as a tool for screening decreased the incidence and mortality of cervical cancer dramatically. This proof of principle study aimed to develop a potential tool for cervical screening using a test that can be applied by patients without visiting a physician and to increase the coverage rate, especially of the high-risk population with low socioeconomic status.

Experimental Design: Human papillomavirus (HPV) DNA testing and methylation analysis of DNA obtained from cervicovaginal specimens of 13, 31, and 11 patients with no dysplasia/low-grade squamous intraepithelial lesion (SIL), high-grade SIL, and invasive cervical cancer, respectively, collected on a tampon, was performed using PCR-based methods to detect invasive cervical cancer and study whether these changes are already present in the precursor lesions.

Results: High-risk HPV DNA was present in 68 and 82% of patients with high-grade SIL and invasive cervical cancer. DNA methylation of the 11 genes tested increased with severity of the cervical lesion. Unsupervised hierarchical cluster analysis using solely information on DNA methylation of the 11 genes was able to predict the presence of invasive cervical cancers: one of the two clusters formed contained 9 of 11 invasive cervical cancers, as well as two high-grade SILs.

Conclusions: HPV DNA and DNA methylation analyzed in cervicovaginal specimens are able to predict invasive cervical cancers. To detect all high-grade SILs when applying this test, genes that become methylated earlier throughout cervical carcinogenesis have to be defined.
Serum Macrophage Inhibitory Cytokine 1 as a Marker of Pancreatic and Other Periampullary Cancers
Authors: Koopmann Jens , Buckhaults Phillip , Brown David A. , Zahurak Marianna L. , Sato Norihiro , Fukushima Noriyoshi , Sokoll Lori J. , Chan Daniel W. , Yeo Charles J. , Hruban Ralph H. , Breit Samuel N. , Kinzler Kenneth W. , Vogelstein Bert , Goggins Michael
Published: September 2005

Purpose: Patients with pancreatic ductal adenocarcinoma usually present with advanced-stage disease and a dismal prognosis. One effective strategy likely to improve the morbidity and mortality from pancreatic cancer would be the identification of accurate, noninvasive diagnostic markers that would enable earlier diagnosis of symptomatic patients and earlier detection of cancer in asymptomatic individuals at high risk for developing pancreatic cancer. In this study, we evaluated serum macrophage inhibitory cytokine-1 (MIC-1) as a marker of pancreatic cancer.

Experimental Design: MIC-1 expression in primary pancreatic cancers, intraductal papillary mucinous neoplasms, and pancreatic cancer cell lines was determined using the National Center for Biotechnology Information serial analysis of gene expression database, oligonucleotide microarrays analysis, in situ hybridization, and immunohistochemistry. Serum MIC-1 levels were determined by ELISA in 80 patients with pancreatic adenocarcinomas, in 30 patients with ampullary and cholangiocellular carcinomas, in 42 patients with benign pancreatic tumors, in 76 patients with chronic pancreatitis, and in 97 healthy control subjects. The diagnostic performance of serum MIC-1 as a marker of pancreatic cancer was compared with that of serum CA19–9.

Results: Oligonucleotide microarray and serial analysis of gene expression data demonstrated that MIC-1 RNA levels were higher in primary pancreatic cancers, intraductal papillary mucinous neoplasms, and pancreatic cancer cell lines than in nonneoplastic pancreatic ductal epithelium. MIC-1 expression was localized to the malignant epithelium in pancreatic adenocarcinomas by in situ hybridization. MIC-1 protein was expressed in 14 of 16 primary pancreatic adenocarcinomas (88%) by immunohistochemistry and was also expressed in some pancreata affected by pancreatitis but not in normal pancreas. Serum MIC-1 levels were significantly higher in patients with pancreatic ductal adenocarcinoma (mean ± SD, 2428 ± 2324 pg/ml) and in patients with ampullary and cholangiocellular carcinomas (2123 ± 2387 pg/ml) than in those with benign pancreatic neoplasms (940 ± 469 pg/ml), chronic pancreatitis (1364 ± 1236 pg/ml), or in healthy controls (546 ± 262 pg/ml). An elevated serum MIC-1 (defined as 2 SD above the mean for healthy controls) performed as well as CA19–9 (area under the receiver operating characteristic curve, 0.81 and 0.77, respectively), and the combination of MIC-1 and CA19–9 significantly improved diagnostic accuracy (P < 0.05; area under the receiver operating characteristic curve, 0.87; sensitivity, 70%; specificity, 85%).

Conclusion: Serum MIC-1 measurement can aid in the diagnosis of pancreatic adenocarcinoma.
NY-ESO-1 Protein Formulated in ISCOMATRIX Adjuvant Is a Potent Anticancer Vaccine Inducing Both Humoral and CD8+ T-Cell-Mediated Immunity and Protection against NY-ESO-1+ Tumors
Authors: Maraskovsky Eugene , Sjölander Sigrid , Drane Debbie P. , Schnurr Max , Le Thuy T. T. , Mateo Luis , Luft Thomas , Masterman Kelly-Anne , Tai Tsin-Yee , Chen Qiyuan , Green Simon , Sjölander Anders , Pearse Martin J. , Lemonnier Francois A. , Chen Weisan , Cebon Jonathan , Suhrbier Andreas
Published: September 2005

NY-ESO-1 is a 180 amino-acid human tumor antigen expressed by many different tumor types and belongs to the family of “cancer-testis” antigens. In humans, NY-ESO-1 is one of the most immunogenic tumor antigens and NY-ESO-1 peptides have been shown to induce NY-ESO-1-specific CD8+ CTLs capable of altering the natural course of NY-ESO-1-expressing tumors in cancer patients. Here we describe the preclinical immunogenicity and efficacy of NY-ESO-1 protein formulated with the ISCOMATRIX adjuvant (NY-ESO-1 vaccine). In vitro, the NY-ESO-1 vaccine was readily taken up by human monocyte-derived dendritic cells, and on maturation, these human monocyte-derived dendritic cells efficiently cross-presented HLA-A2-restricted epitopes to NY-ESO-1-specific CD8+ T cells. In addition, epitopes of NY-ESO-1 protein were also presented on MHC class II molecules to NY-ESO-1-specific CD4+ T cells. The NY-ESO-1 vaccine induced strong NY-ESO-1-specific IFN-γ and IgG2a responses in C57BL/6 mice. Furthermore, the NY-ESO-1 vaccine induced NY-ESO-1-specific CD8+ CTLs in HLA-A2 transgenic mice that were capable of lysing human HLA-A2+ NY-ESO-1+ tumor cells. Finally, C57BL/6 mice, immunized with the NY-ESO-1 vaccine, were protected against challenge with a B16 melanoma cell line expressing NY-ESO-1. These data illustrate that the NY-ESO-1 vaccine represents a potent therapeutic anticancer vaccine.
The Clinical Application of Targeting Cancer through Histone Acetylation and Hypomethylation
Authors: Gilbert Jill , Gore Steve D. , Herman James G. , Carducci Michael A.
Published: September 2005

Methods of gene inactivation include genetic events such as mutations or deletions. Epigenetic changes, heritable traits that are mediated by changes in DNA other than nucleotide sequences, play an important role in gene expression. Two epigenetic events that have been associated with transcriptional silencing include methylation of CpG islands located in gene promoter regions of cancer cells and changes in chromatin conformation involving histone acetylation. Recent evidence demonstrates that these processes form layers of epigenetic silencing. Reversal of these epigenetic processes and up-regulation of genes important to prevent or reverse the malignant phenotype has therefore become a new therapeutic target in cancer treatment.
Predicting Radiosensitivity Using DNA End-Binding Complex Analysis
Authors: Ismail Sheikh M. , Puppi Monica , Prithivirajsingh Sheela , Munshi Anupama , Raju Uma , Meyn Raymond E. , Buchholz Thomas A. , Story Michael D. , Brock William A. , Milas Luka , Thames Howard D. , Stevens Craig W.
Published: September 2005

Previous reports have suggested that measuring radiosensitivity of normal and tumor cells would have significant clinical relevance for the practice of radiation oncology. We hypothesized that radiosensitivity might be predicted by analyzing DNA end-binding complexes (DNA-EBCs), which form at DNA double-strand breaks, the most important cytotoxic lesion caused by radiation. To test this hypothesis, the DNA-EBC pattern of 21 primary human fibroblast cultures and 15 tumor cell lines were studied. DNA-EBC patterns were determined using a modified electrophoretic mobility shift assay and were correlated with radiosensitivity, as measured by SF2. DNA-EBC analysis identified a rapidly migrating ATM-containing band (identified as “band-A”) of which the density correlated with SF2 (0.02 ≤ SF2 ≤ 0.41) in primary fibroblasts (r2 = 0.77). The DNA-EBC pattern of peripheral blood lymphocytes was identical to that of fibroblasts. In addition, band-A density correlated with SF2 (0.35 ≤ SF2 ≤ 0.80) in 15 human tumor cell lines (r2 = 0.91). Densitometry of other bands, or total DNA-EBC binding, correlated more poorly with SF2 (r2 < 0.45). These data indicate that DNA-EBC analysis may be a practical, clinically relevant predictor of tumor and primary cell radiosensitivity.
Molecular Differences between Sporadic Serrated and Conventional Colorectal Adenomas
Authors: Konishi Kazuo , Yamochi Toshiko , Makino Reiko , Kaneko Kazuhiro , Yamamoto Taikan , Nozawa Hisako , Katagiri Atsushi , Ito Hiroaki , Nakayama Kentarou , Ota Hidekazu , Mitamura Keiji , Imawari Michio
Published: September 2005

Purpose: The purpose is to compare the molecular characteristics of serrated adenomas (SAs) with those of conventional adenomas (CADs) and hyperplastic polyps (HPs).

Experimental Design: We evaluated the proliferative activity and molecular alterations in 47 SAs (25 pure-type and 22 mixed-type), 71 CADs, and 23 HPs.

Results: The proliferative activity of SAs, as evaluated by Ki-67 expression, was intermediate between CADs and HPs. There was no significant difference in the incidence of KRAS or p53 mutations between the three histological groups. In the microsatellite instability (MSI) analysis, 21% of SAs (9 of 43) showed MSI at two or more loci (MSI-H); corresponding values were 5% of CADs (3 of 64) and 8% of HPs (1 of 13; SAs versus CADs, P = 0.0125). MSI-H was more likely to be found in pure-type SAs (36%; 8 of 22) than in mixed-type SAs (5%; 1 of 21; P = 0.0212). Loss of hMLH-1 expression was found in 8 of 9 SAs with MSI-H. The incidence of BRAF or KRAS mutations was 36 and 15% of SAs, respectively; the combined incidence of BRAF and KRAS mutations occurred in 49% of SAs. However, there was no significant difference in the incidence of BRAF or KRAS mutations between SAs with and without MSI-H.

Conclusions: Genetic instability is more frequently implicated in the tumorigenesis of SAs, especially pure-type SAs, than in that of CADs. In contrast, activation of the Ras/Raf/MEK/MAP kinase cascade by BRAF or KRAS mutation, independently of the genetic instability, may be associated with the progression of about half of SAs.
Enhanced Chemosensitivity to Irinotecan by RNA Interference-Mediated Down-Regulation of the Nuclear Factor-κB p65 Subunit
Authors: Guo Jun , Verma Udit N. , Gaynor Richard B. , Frenkel Eugene P. , Becerra Carlos R.
Published: September 2005

In preclinical tumor models, inhibition of nuclear factor-κB (NF-κB) has been associated with increased sensitivity to chemotherapeutic agents such as irinotecan (CPT-11). This is based on the fact that a variety of chemotherapy agents such as CPT-11 activate NF-κB to result in the expression of genes such as c-IAP1 and c-IAP2 that might be responsible for the inhibition of chemotherapy-induced apoptosis. In this study, RNA interference [small interfering RNA (siRNA)] was used to down-regulate the NF-κB p65 subunit in the HCT116 colon cancer cell line, and its role, in the presence and absence of CPT-11, was assessed on cell growth and apoptosis. Reduction of endogenous p65 by siRNA treatment significantly impaired CPT-11-mediated NF-κB activation, enhanced apoptosis, and reduced colony formation in soft agar. Furthermore, the in vivo administration of p65 siRNA reduced HCT116 tumor formation in xenograft models in the presence but not the absence of CPT-11 administration. These data indicate that the administration of siRNA directed against the p65 subunit of NF-κB can effectively enhance in vitro and in vivo sensitivity to chemotherapeutic agents.
Phase I Study of the Farnesyltransferase Inhibitor Lonafarnib with Paclitaxel in Solid Tumors
Authors: Khuri Fadlo R. , Glisson Bonnie S. , Kim Edward S. , Statkevich Paul , Thall Peter F. , Meyers Michael L. , Herbst Roy S. , Munden Reginald F. , Tendler Craig , Zhu Yali , Bangert Sandra , Thompson Elizabeth , Lu Charles , Wang Xue-Mei , Shin Dong M. , Kies Merrill S. , Papadimitrakopoulou Vali , Fossella Frank V. , Kirschmeier Paul , Bishop W. Robert , Hong Waun Ki
Published: September 2005

Purpose: To establish the maximum tolerated dose of lonafarnib, a novel farnesyltransferase inhibitor, in combination with paclitaxel in patients with solid tumors and to characterize the safety, tolerability, dose-limiting toxicity, and pharmacokinetics of this combination regimen.

Experimental Design: In a Phase I trial, lonafarnib was administered p.o., twice daily (b.i.d.) on continuously scheduled doses of 100 mg, 125 mg, and 150 mg in combination with i.v. paclitaxel at doses of 135 mg/m2 or 175 mg/m2 administered over 3 h on day 8 of every 21-day cycle. Plasma paclitaxel and lonafarnib concentrations were collected at selected time points from each patient.

Results: Twenty-four patients were enrolled; 21 patients were evaluable. The principal grade 3/4 toxicity was diarrhea (5 of 21 patients), which was most likely due to lonafarnib. dose-limiting toxicities included grade 3 hyperbilirubinemia at dose level 3 (100 mg b.i.d. lonafarnib and 175 mg/m2 paclitaxel); grade 4 diarrhea and grade 3 peripheral neuropathy at dose level 3A (125 mg b.i.d. lonafarnib and 175 mg/m2 paclitaxel); and grade 4 neutropenia with fever and grade 4 diarrhea at level 4 (150 mg b.i.d. lonafarnib and 175 mg/m2 paclitaxel). The maximum tolerated dose established by the continual reassessment method was lonafarnib 100 mg b.i.d. and paclitaxel 175 mg/m2. Paclitaxel appeared to have no effect on the pharmacokinetics of lonafarnib. The median duration of therapy was eight cycles, including seven cycles with paclitaxel. Six of 15 previously treated patients had a durable partial response, including 3 patients who had previous taxane therapy. Notably, two of five patients with taxane-resistant metastatic non-small cell lung cancer had partial responses.

Conclusions: When combined with paclitaxel, the recommended dose of lonafarnib for Phase II trials is 100 mg p.o. twice daily with 175 mg/m2 of paclitaxel i.v. every 3 weeks. Additional studies of lonafarnib in combination regimens appear warranted, particularly in patients with non-small cell lung cancer.
Skp2
Gene Copy Number Aberrations Are Common in Non-Small Cell Lung Carcinoma, and Its Overexpression in Tumors with
ras
Mutation Is a Poor Prognostic Marker
Authors: Zhu Chang Qi , Blackhall Fiona H. , Pintilie Melania , Iyengar Pratibha , Liu Ni , Ho James , Chomiak Taylor , Lau Davina , Winton Timothy , Shepherd Frances A. , Tsao Ming-Sound
Published: September 2005

Purpose: Skp2 plays a critical role in cell cycle progression, especially at the G1-S transition, putatively through its control of several cell cycle regulator proteins. The Skp2 gene is located on a region of chromosome 5p that is commonly overrepresented in lung cancer. The present study aimed to evaluate Skp2 abnormalities and their prognostic value in non-small cell lung cancer (NSCLC).

Experimental Design: In total 16 NSCLC cell lines and 163 primary tumors were included in studies to measure Skp2 relative gene copy number, mRNA abundance, and protein level. The tumors were also evaluated for p27 protein expression level and ras mutation. These values were correlated with the clinical and pathological features of the patients.

Results: Skp2 relative gene copy number aberrations were found in 88 and 65% of NSCLC cell lines and primary tumors, respectively. Overrepresentation was especially common among squamous cell carcinoma (74%). Both gene copy overrepresentation (13%) and loss (35%) were found in adenocarcinoma. Skp2 relative gene copy number was significantly correlated with mRNA and protein levels, but none of these were correlated with p27 protein levels. Neither high Skp2 protein expression nor ras mutation was prognostically significant. In NSCLCs with ras mutation, however, high Skp2 protein expression was a significant independent poor prognostic marker.

Conclusion: There appears to be a synergistic interaction between high Skp2 protein expression and ras mutation with negative impact on the survival of NSCLC patients.
Resistance to Tyrosine Kinase Inhibition by Mutant Epidermal Growth Factor Receptor Variant III Contributes to the Neoplastic Phenotype of Glioblastoma Multiforme
Authors: Learn Chris A. , Hartzell Tristan L. , Wikstrand Carol J. , Archer Gary E. , Rich Jeremy N. , Friedman Allan H. , Friedman Henry S. , Bigner Darell D. , Sampson John H.
Published: September 2005

Purpose: We have reported previously that tumors expressing wild-type epidermal growth factor receptor (EGFR) in a murine model are sensitive to the EGFR tyrosine kinase inhibitor gefitinib, whereas tumors expressing mutant EGFR variant III (EGFRvIII) are resistant. Determination of how this differential inhibition occurs may be important to patient selection and treatment criteria, as well as the design of future therapeutics for glioblastoma multiforme.

Experimental Design: We have determined and quantified how treatment with gefitinib at commonly used, noncytotoxic doses affects neoplastic functions ascribed to EGFRvIII, including downstream signaling by Akt, DNA synthesis, and cellular invasion. In doing so, we have tested and compared a series of wild-type and mutant EGFRvIII-expressing fibroblast and glioblastoma cell lines in vitro after treatment with gefitinib.

Results: The results of these experiments demonstrate that short-term treatment with gefitinib (∼24 h) does not reduce phosphorylation of EGFRvIII, whereas EGFR phosphorylation is inhibited in a dose-dependent manner. However, after daily treatment with gefitinib, phosphorylation declines for EGFRvIII by day 3 and later. Nevertheless, after 7 days of daily treatment, cells that express and are dependent on EGFRvIII for tumorigenic growth are not effectively growth inhibited. This may be due in part to phosphorylation of Akt, which is inhibited in EGFR-expressing cells after treatment with gefitinib, but is unaffected in cells expressing EGFRvIII. Cell cycle analysis shows that nascent DNA synthesis in EGFR-expressing cells is inhibited in a dose-dependent manner by gefitinib, yet is unaffected in EGFRvIII-expressing cells with increasing dosage. Furthermore, cells expressing EGFRvIII demonstrate greater invasive capability with increasing gefitinib concentration when compared with cells expressing EGFR after treatment.

Conclusions: We conclude that the neoplastic phenotype of EGFRvIII is relatively resistant to gefitinib and requires higher doses, repeated dosing, and longer exposure to decrease receptor phosphorylation. However, this decrease does not effectively inhibit the biologically relevant processes of DNA synthesis, cellular growth, and invasion in cells expressing EGFRvIII.