Cancer-free Patients Research Articles

Metabolic Syndrome Is Also a Risk Factor for Primary Liver Cancer in Patients Younger than 65 Years of Age?

We read with great interest the article by Welzel et al.1 The study reported a positive association between metabolic syndrome and hepatocellular carcinoma (HCC), but the results were restricted to people aged ≥65 years of age. Although the incidence of obesity and/or metabolic syndrome is showing no sign of decline, the epidemic of hepatitis B (HBV) and C (HCV) viruses is at its peak, thus increasing the incidence of HCC.2, 3 To provide further insights on the association of HCC and metabolic syndrome on the interaction with HBV/HCV infection, we report data from an Italian case–control study on HCC. The study was conducted between 1999 and 2002 in the province of Pordenone, Northeastern Italy, and in Naples, Southern Italy.4 Cases comprised 185 patients aged 43-84 years (50% below age 65 years) with incident HCC, who had not yet received any cancer treatment at study entry. The control group included 412 cancer-free patients from the same areas as the cases.4 Metabolic syndrome was associated with a four-fold higher risk of HCC. The association was confirmed among hepatitis B surface antigen (HBsAg)-negative and antibody to HCV (anti-HCV)-negative subjects (odds ratio [OR] = 4.00; 95% confidence interval [CI] = 1.30-12.27). According to the individual preexisting medical condition, only diabetes was significantly associated with HCC risk (OR = 3.75; 95% CI = 1.66-8.44; Table 1). This result was in agreement with a recent study.5 Our results are therefore in agreement with the study by Welzel et al.1; in particular, our study confirms that metabolic syndrome is also a risk factor for HCC in people younger than 65 years (50% of our study population). Our results on metabolic syndrome, together with heavy drinking (in the North) and high HCV prevalence (in the South), contribute to explain the high incidence of and mortality rates from HCC in Italy.3, 6 In order to reduce the burden of liver cancer, actions to control the recent epidemic of metabolic syndrome should be promoted. The authors gratefully acknowledge The Italian League Against Cancer (LILT), Project n. 11/2008. Maurizio Montella M.D.*, Jerry Polesel Sc.D. , Renato Talamini M.D. , Anna Crispo Sc.D.*, Aldo Giudice Sc.D.*, Francesco Izzo M.D. , Carlo La Vecchia Ph.D.§ ¶, * Unità di Epidemiologia, stituto Nazionale dei Tumori Fondazione “G. Pascale” Naples, Italy, Dipartimento di Epidemiologia e Biostatistica, Centro di Riferimento Oncologico—IRCCS, Aviano, Italy, Divisione di Chirurgia “D”, Istituto Nazionale dei Tumori Fondazione “G. Pascale,” Naples, Italy, § Istituto di Ricerche Farmacologiche “Mario Negri,” Milan, Italy, ¶ Istituto di Statistica Medica e Biometria “G. A. Maccacaro,” Università degli Studi di Milano, Milan, Italy.

The Importance of Being Negative

The Importance of Being Negative

No association between angiotensin I converting enzyme (ACE) I/D polymorphism and gastric cancer risk among Japanese.

The angiotensin I-converting enzyme (ACE) is involved in cell proliferation, angiogenesis, inflammation, and tissue remodeling, all of which could play a role in carcinogenesis. The DD genotype of ACE I/D polymorphism with a higher ACE level than either ID or II genotypes was reported to increase the risk of several cancers. This is a case-control study examining the association between the polymorphism and gastric cancer risks among Japanese. Cases numbered 583 patients aged 27 to 80 years with gastric cancer diagnosed at the Aichi Cancer Center Hospital from 2001 to 2005. Controls were 1,742 sex and age frequency-matched cancer-free patients, who visited the same hospital during that same period. The ACE I/D polymorphism was genotyped using a polymerase chain reaction with confronting two-pair primers. The results showed that the age- and sex- adjusted ORs of gastric cancer were 0.95 (95% CI, 0.78-1.16) for ID, and 1.08 (95% CI, 0.80-1.46) for DD relative to II. Among the controls with H. pylori sero-positive or gastric atrophy (GA), the ORs of ID and DD relative to II were 1.20 (95% CI, 0.88-1.63) and 1.16 (95% CI, 0.73-1.84) for mild GA, and 1.22 (95% CI, 0.84-1.78) and 1.08 (95% CI, 0.61-1.89) for severe GA, respectively. In conclusion, there was no significant association of the ACE I/D polymorphism with the risk of gastric cancer. Among the controls, the polymorphism was not associated with the severity of GA.

Highlights from Recent Cancer Literature

Highlights from Recent Cancer Literature

Malignant Disease Within 5 Years After Surgery in Relation to Duration of Sevoflurane Anesthesia and Time with Bispectral Index Under 45

Surgery, general anesthesia, and related events have been implicated to promote cancer proliferation. We investigated the incidence of cancer within 5 years after surgery in relation to duration of anesthesia (T(ANESTH)) and also by time with bispectral index (BIS) under 45 (T(BIS<45)) serving as a proxy for more profound anesthesia exposure. New malignant diagnoses after surgery under sevoflurane anesthesia were obtained in a prospective cohort of 2972 BIS-monitored patients without any clinically diagnosed malignant disease at the time of index surgery. The risk of cancer during follow-up in relation to T(ANESTH) and T(BIS<45) was assessed by Cox regression. The cancer incidence in this surgical population was compared with the incidence in a standardized general population by calculation of standard incidence ratio. One hundred twenty-nine patients (4.3%) were assigned 136 new malignant diagnoses within 5 years after surgery. No relation between T(ANESTH) or T(BIS<45) and new malignant disease was found, nor were any significant relations obtained when other thresholds for BIS (i.e., <30, <40, and <50, respectively) were used in the calculations. The standard incidence ratio for new malignant disease was 1.37 (confidence interval, 1.15-1.62). Neither duration of anesthesia nor increased cumulative time with profound sevoflurane anesthesia was associated with an increased risk for new malignant disease within 5 years after surgery in previously cancer-free patients.

Abstract 1161: Detection and identification of a miRNA expression profile from cell-free urine: Potential utility in bladder cancer

Abstract Introduction/Objective: MicroRNAs are small, non-coding RNAs that have been shown to play an important role in tumorigenesis. There is differential expression of miRNA in cancer progression, and profiling of miRNA is promising for both diagnosis and treatment of malignant tumors. In this study we isolated RNA from cell-free urine in an attempt to characterize miRNA profiles indicating the presence of urothelial carcinoma and its potential use as a non-invasive assay to identify patients with cancer progression. Methods: Urine was collected from patients diagnosed with bladder cancer and control patients with no history of cancer under an IRB-approved protocol. Urine was centrifuged and total RNA was isolated from the supernatants using the mirVana Paris™ kit. A total of 178 samples were grouped according to grade and stage (healthy controls (35), TaG1 (19), T1G3 (16), ≥T2 (30), carcinoma in situ (CIS; 28) and no evidence of disease following therapy (50). Seven hundred and thirty miRNAs were profiled by qRT-PCR on pooled samples within each group. Validation of selected miRNAs was performed on individual samples using qRT-PCR. Results: Cell-free RNA was isolated from urine of 35 healthy controls and 143 patients with bladder cancer. Of the 730 miRNAs tested, 236 were detected in at least one of the pooled samples using a Ct cutoff of 35. The number of miRNAs detected in the pooled samples correlated with disease progression where the healthy control group and the ≥T2 group expressed 8 and 228 miRNAs, respectively. qRT-PCR of individual samples revealed a gradual increase of some miRNAs with disease progression. Statistical analysis adjusted for multiple comparisons demonstrated differences between groups based on miRNA expression levels. In addition, a panel of miRNAs was identified which discriminated between cancer and cancer-free patients. Conclusion: This study demonstrates the successful isolation of miRNAs from cell-free urine. Utilizing non-invasive urine based assays, we identified a miRNA panel that can discriminate between cancer-free patients and patients with urinary carcinoma of the bladder. These findings provide evidence that profiling of miRNAs from cell-free urine holds the promise for the development of valuable clinical tools. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1161. doi:10.1158/1538-7445.AM2011-1161

Tobacco smoking, alcohol drinking, and the risk of different histological types of nasopharyngeal cancer in a low-risk population

Nasopharyngeal carcinoma (NPC) is rare in western Countries. Tobacco smoking is a well-recognised risk factor, whereas the role of alcohol drinking is still in debate. We conducted a hospital-based case-control study in Italy on 150, histologically-confirmed, NPC cases of Caucasian ethnicity, aged 18-76years, including 118 undifferentiated NPCs and 22 differentiated squamous-cell NPC. Controls were 450 Caucasian cancer-free patients admitted to general hospitals for acute conditions. Cases and controls were matched according to sex, age, and place of residence. Logistic regression was used to estimate odds ratios (OR) and the corresponding 95% confidence intervals (CI) while adjusting for known confounders. No significant association emerged between tobacco smoking and all NPCs (OR for current vs. never smokers=1.52; 95% CI: 0.89-2.60). Conversely, for differentiated NPC only, statistically significant elevated OR were associated with increasing smoking intensity (OR for ⩾15cigarettes/day=5.40; 95% CI: 1.34-21.76) and duration of the habit (OR for ⩾32years=4.48; 95% CI: 1.11-18.04). Although alcohol drinking was not, per se, significantly associated to NPC risk, the combination of tobacco smoking and alcohol drinking accounted for 57% of differentiated NPCs, whereas it accounted for only 14% of undifferentiated carcinomas. Our findings suggest that, in western populations, NPC includes two separate entities: the differentiated NPC, associated with tobacco smoking like other cancers of head and neck, and the undifferentiated NPC, upon which tobacco smoking has little or no influence.

1362 DETECTION AND IDENTIFICATION OF A MIRNA EXPRESSION PROFILE FROM CELL-FREE URINE: POTENTIAL UTILITY IN BLADDER CANCER

INTRODUCTION AND OBJECTIVES: MicroRNAs are small, non-coding RNAs that have been shown to play an important role in tumorigenesis. There is differential expression of miRNA in cancer progression, and profiling of miRNA is promising for both diagnosis and treatment of malignant tumors. In this study we isolated RNA from cell-free urine in an attempt to characterize miRNA profiles indicating the presence of urothelial carcinoma and its potential use as a noninvasive assay to identify patients with cancer progression. METHODS: Urine was collected from patients diagnosed with bladder cancer and control patients with no history of cancer under an IRB-approved protocol. Specimens were then centrifuged and total RNA was isolated from the supernatants using the mirVana ParisTM kit. A total of 178 samples were grouped according to grade and stage: healthy controls (35), TaG1 (19), T1G3 (16), T2 (30), carcinoma in situ (CIS; 28) and no evidence of disease following therapy (50). Seven hundred and thirty miRNAs were profiled by qRT-PCR on pooled samples within each group. Validation of selected miRNAs was performed on individual samples using qRT-PCR. RESULTS: Cell-free RNA was isolated from urine of 35 healthy controls and 143 patients with bladder cancer. Of the 730 miRNAs tested, 236 were detected in at least one of the pooled samples using a cycle threshold cutoff of 35. The number of miRNAs detected in the pooled samples correlated with disease progression where the healthy control group and the T2 group expressed 8 and 228 miRNAs, respectively. qRT-PCR of individual samples revealed a gradual increase of some miRNAs with disease progression. Statistical analysis adjusted for multiple comparisons demonstrated differences between groups based on miRNA expression levels. In addition, a panel of miRNAs was identified which discriminated between cancer and cancer-free patients. CONCLUSIONS: This study demonstrates the successful isolation of miRNAs from cell-free urine. Utilizing non-invasive urine based assays, we identified a miRNA panel that can discriminate cancer-free patients and patients with urinary carcinoma of the bladder. These findings provide evidence that profiling of miRNAs from cell-free urine holds the promise for the development of valuable clinical tools.

Clinical study of autoantibody spectrum against ovarian cancer associated antigens combined with CA(125) in detecting and monitoring ovarian cancer

To evaluate the clinical value of autoantibody spectrum against ovarian cancer associated antigens combine CA(125) in detecting and monitoring ovarian cancer. Circulating IgG, IgM autoantibodies against ovarian cancer associated antigens which included TM4SF1, C1D, TIZ, OV-142, FXR1 and OV-189 were measured by indirect ELISA in serum from 126 patients with ovarian cancer (prior treatment), 42 patients with benign ovarian masses, 142 healthy women. Cut off value of IgG, IgM autoantibodies were determined by receive operating characteristic (ROC) curve. CA(125) was measured in serum by immunoradiometric assay (IRMA). We evaluated the clinical value of combining multiple autoantibodies (autoantibody spectrum), combining autoantibody spectrum with CA(125) by binary logistic regresion. The positive ratio of autoantibody spectrum in serum (prior and post treatment) of 24 synchronization patients with ovarian cancer was analyzed to evaluate the value in monitoring state of illness. Our data indicated that serum contains IgG, IgM autoantibodies against ovarian cancer associated antigens. The positive ratio of IgG autoantibodies in serum from ovarian cancer patients and cancer-free patients were 34.1% - 47.6% and 13.0% - 19.0%, respectively (P < 0.05). The positive ratio of IgM autoantibodies in serum from ovarian cancer patients and cancer-free patients were 39.7% - 53.2% and 12.0% - 33.2%, respectively (P < 0.05). The positive ratio of IgG autoantibodies against FXR1 and IgM autoantibodies against TIZ, FXR1 and OV-189 in early stage (I-II) ovarian cancer (55.3%, 63.8%, 61.7% and 66.0%) were significantly higher than those in advanced (III-IV) ovarian cancer (34.2%, 39.2%, 26.6%, 45.6%; all P < 0.05). Combining five autoantibodies (TM4SF1 IgG, TM4SF1 IgM, C1D IgG, FXR1 IgG and TIZ IgM) showed significantly improved sensitivity (75.4%, P < 0.05), lower specificity (78.3%, P < 0.05) and similar accuracy (77.1%, P > 0.05) in detecting ovarian cancer compared to those of CA(125) (61.1%, 88.0%, 77.1%). But the autoantibody spectrum showed significantly improved sensitivity in classifying early stage (76.6%), compared to those of CA(125) (51.1%, P < 0.05). Combining autoantibody spectrum with CA(125) showed significantly improved sensitivity (85.7%), specificity (90.8%)and accuracy (88.7%) in detecting ovarian cancer compared to those of autoantibody spectrum alone (all P < 0.05), while CA(125) (61.1%, P < 0.05; 88.0%, P > 0.05; 77.1%, P < 0.05). The positive ratio of combine the autoantibody spectrum with CA(125) was significantly lower in 24 post-treatment serum (42%) compared to the pairing prior treatment serum (88%, P < 0.05). Combining the autoantibody spectrum against ovarian cancer associated antigens with CA(125) can improve sensitivity, specificity and accuracy in detecting early ovarian cancer and may be used to monitoring state of illness.

Sporadic breast cancer patients' germline DNA exhibit an AT‐rich microsatellite signature

Using a custom CGH-like oligonucleotide array to measure the global microsatellite content in the genomes of 72 cancer, cancer-free, and high risk patient and cell line samples (56 germline DNA and 16 in tumor or tumor cell line DNA) we found a unique, reproducible, and statistically significant pattern of 18 motif-specific microsatellite families (out of 962 possible 1-6 mer repeats) in breast cancer patient germline and tumor DNA, but not in germline DNA of cancer-free volunteer controls or in breast cancer patients with BRCA1/2 mutations. These high-similarity A/T rich repetitive motifs were also more pronounced in the germlines and tumors of colon cancer tumor patients (3/6 samples) and microsatellite unstable colon cancer cell lines; however, germline DNA of sporadic breast cancer patients exhibited the largest global content shift for those motifs with extreme AT/GC ratios. These results indicate that global microsatellite variability is complex, suggest the existence of a previously unknown genomic destabilization mechanism in breast cancer patients' germline DNA, and warrant further testing of such microsatellite variability as a predictor of future breast cancer development.

IMMUNOCHEMICAL IDENTIFICATION OF AN EPITHELIAL MEMBRANE ANTIGEN IN PATIENTS WITH BLADDER CANCER

An epithelial membrane antigen (EMA) has been analyzed with Western blot at 130-kDa. ELISA was used for screening and diagnosis of EMA in urine samples of bladder cancer patients. EMA was detected in 43 out of 51 bladder cancer patients with detection rate 84%, while it was detected in 22 out of 28 suspicious patients with detection rate 78.5%. In addition, EMA was detected in 21 out of 51 bladder cancer-free patients with detection rate 41%. Moreover EMA was detected in 1 out of 32 urine samples from healthy individuals with detection rate 3.1%. The performance characteristics of ELISA as a rapid diagnostic assay of bladder carcinoma based on EMA detection in urine samples revealed that the sensitivity was 84%, while the specificity was 96.9% among normal individuals. There was an extremely significant (P 0.0001) higher than its level in bladder cancer-free patients. EMA levels were progressively increased with grades and stages of bladder cancer. It is concluded that, EMA detected at 130 kDa using Western blot and assayed using ELISA for screening and diagnosis of bladder cancer can be used with high sensitivity compared to cystoscopy and high specificity among healthy individuals. Also quantitative estimation of EMA level in the urine of bladder cancer patients can differentiate them from other bladder affections. Therefore, it is concluded that urine EMA assay may serve as a marker for bladder cancer assessment.

Addition of transcatheter arterial chemoembolization decreased local recurrence but had no survival benefit to percutaneous ethanol injection therapy for patients with small hepatocellular carcinoma: A multicenter randomized control study

To assess the efficacy of the additional treatment of transcatheter arterial chemoembolization (TACE) to percutaneous ethanol injection (PEI) therapy for relatively small hepatocellular carcinomas (HCCs), a multicenter randomized control study (RCT) was performed. We conducted an RCT and follow-up study during the enrollment period from 1997 to 1999. Newly diagnosed patients with one to three HCC tumors measuring from 2 to 4 cm (4 cm maximum) in diameter were enrolled. A total of 30 patients initially underwent a combination TACE-PEI or PEI-alone therapies at eight randomly assigned Japanese hospitals. However, 3 patients withdrew. Of the 27 remaining patients, 13 were treated with the combination TACE-PEI therapy and 14 with PEI therapy alone. The patients were observed over several months [median (interquartile range) 33.2 (24.6) months]. There were no significant differences in the background of the patients between the two groups. Among the patients treated with TACE-PEI, the development of a local residual tumor was of significantly lower occurence, compared to the group receiving PEI alone (7.6 and 42.9%, respectively; P=0.024). However, the mean cancer-free time (absence of local or multiple nodule recurrence) or patient survival time was not significantly different between the two groups [PEI alone vs. TACE-PEI: cancer-free time 16.7 (95% CI 7.3-26.0) vs. 22.9 months (95% CI 12.4-33.4); survival time 57.2 (95% CI 37.2-77.2) vs. 42.4 months (95% CI 29.2-55.6)]. Although the combination of TACE and PEI had significant effects on the local tumor control, no efficacy of the addition of TACE to PEI was noted in the prognosis among patients with relatively small HCC tumors.

Fragment c gamma receptor gene polymorphisms and breast cancer risk in case–control studies in Japanese, Japanese Brazilians, and non-Japanese Brazilians

Previous studies showing the presence of antibodies against tumor-associated antigens in healthy individuals suggest that antibody-dependent cell cytotoxicity (ADCC) might play a role in the development of breast cancer. We hypothesized that functional polymorphisms in fragment c gamma receptor (FcgR) genes were associated with breast cancer risk. We conducted hospital-based case-control studies of patients aged 20-74 years with invasive breast cancer, and matched controls from medical checkup examinees in Nagano, Japan and from cancer-free patients in São Paulo, Brazil. A total of 869 pairs (403 Japanese, 80 Japanese Brazilians and 386 non-Japanese Brazilians) were genotyped for two single nucleotide polymorphisms (SNPs): a histidine (H)/arginine (R) polymorphism at position 131 of FcgRIIa (FcgRIIa H131R) and a valine (V)/phenylalanine (F) polymorphism at position 158 of FcgRIIIa (FcgRIIIa F158V). We found no statistically significant association between either of the two SNPs and breast cancer risk regardless of population. In analyses of the three populations combined, adjusted odds ratio (OR) was 0.93 [95% confidence interval (CI) 0.66-1.32] for women with the R/R versus H/H genotype of the FcgRIIa H131R polymorphism and 1.04 (95% CI 0.69-1.57) for the V/V versus F/F genotype of the FcgRIIIa F158V polymorphism. On combination of the two SNPs, compared to women with both the R/R genotype of the FcgRIIa H131R polymorphism and F/F genotype of the FcgRIIIa F158V polymorphism, the adjusted OR for women with both the H/H and V/V genotype was 0.68 (95% CI 0.37-1.27). In conclusion, our findings suggest that ADCC might not play a major role in the etiology of breast cancer.

Detection of lung cancer in silicosis patients using a tumor-marker panel

This study evaluates the diagnostic power of a fuzzy classifier and a tumor marker panel for the detection of lung cancers in comparison to pneumoconiosis patients at high-risk of developing lung cancer. CEA, CYFRA 21-1, NSE, SCC and CRP were measured in newly diagnosed lung cancer patients of different histological types and stages in comparison to patients suffering from silicosis. First a fuzzy classifier was generated with a set of 216 primary lung cancer patients (158 non-small cell lung cancer (NSCLC) and 58 small cell lung cancer (SCLC) patients) and 127 patients suffering from pneumoconiosis (51 silicosis and 76 asbestosis). Subsequently, the classifier was validated on a second cohort of 38 NSCLC patients, 55 silicosis patients, 32 patients with chronic obstructive airway diseases (COLD) and 28 healthy control subjects. At 95%-specificity, NSCLC patients were detected in 50% at stage I (n= 30), in 64% at stage II (n=22), in 82% at stage III (n=56), in 88% at stage IV (n=50) and SCLC patients with limited disease status (n=21) in 71% and with extensive disease status (n=37) in 89% by use of the fuzzy classifier. Detection rates of single markers were below those. For the best single marker in NSCLC, CYFRA 21-1, sensitivities were 23.3% at stage I, 45.4% at stage II, 71.4% at stage III, 84% at stage IV (n=50), respectively. For the best single marker in SCLC, NSE, sensitivities were 61.9% at stage of limited disease and 81.1% at stage of extensive disease. In the validation set, the fuzzy classifier showed correct negative classification in 49 of the 55 cancer-free silicosis patients (specificity: 89%), in all COLD patients (specificity: 100%) and in all but one healthy subject (specificity: 96%). This confirmed an overall specificity of 93.9%. The sensitivity for lung cancer detection in high risk populations was 73.6%. All large cell carcinomas could be detected. The positive predictive value was 80%. The negative predictive value reached 91.5%. With the fuzzy classifier and a marker panel a reliable diagnostic tool for the detection of lung cancers in a high risk population is available.

Influence of Surgical Trauma on Neopterin Concentrations in Gastric Carcinoma Patients

Impairment of immune surveillance in the immediate postoperative period may accelerate the growth of tumor cells that remain despite radical resection in advanced cases. The present study was undertaken to evaluate the impact of major surgical trauma on host cellular immunity of gastric carcinoma cases. Sixty-eight consecutive patients with gastric carcinoma were divided into three subgroups according to the classification of the American Joint Committee on Cancer Staging, and they underwent surgery with a curative intent. Thirty-eight cancer-free patients served as controls, and they underwent surgical management for benign diseases. Physiological and Operative Severity Score for the enUmeration of Mortality and Morbidity (POSSUM) risk adjustment system was used to estimate the operative and physiological scores of patients. Plasma cortisol, serum interleukin-6 and high-sensitive C-reactive protein levels were determined in order to assess the severity of trauma-related inflammatory response. The frequency of increased neopterin concentrations of cancer patients was estimated by comparison with the average values of controls. Response of interleukin-6 to surgery was closely related with the postoperative cortisol of cancer patients, but not correlated with neopterin and high-sensitive C-reactive protein levels. Although the extent of tumor invasion might have gradually decreased the macrophage response to surgical trauma, the overall increase in postoperative neopterin levels of cancer cases was highly significant. However, maximal frequency of increased neopterin concentration was obtained in advanced group. Macrophages might recognize the traumatic challenges in every stage of gastric cancer. Increasing individual neopterin concentration was not solely specific for tumor growth, but it was partially predictive of immune competence even in advanced cases.

Genetic variants in selected pre‐microRNA genes and the risk of squamous cell carcinoma of the head and neck

Single nucleotide polymorphisms (SNPs) in microRNAs (miRNAs) may alter the processing, transcription, and expression of miRNAs and, thus, may contribute to cancer development. The authors hypothesized that common polymorphisms in pre-miRNAs are associated individually and (more likely) collectively with the risk of squamous cell carcinoma of the head and neck (SCCHN). The authors genotyped 4 common polymorphisms in pre-miRNAs: Homo sapiens miRNA 146a (hsa-mir-146a) (reference SNP 2910164 [rs2910164]; guanine to cytosine [G→C]), hsa-mir-149 (rs2292832; guanine to thymine [G→T]), hsa-mir-196a2 (rs11614913; C→T), and hsa-mir-499 (rs3746444; adenine to guanine [A→G]) in 1109 patients with SCCHN (cases) and in 1130 cancer-free patients (controls) in a non-Hispanic white population that was frequency-matched by age and sex. Univariate and multivariate logistic regression models were used to calculate crude and adjusted odds ratios (ORs) and 95% confidence intervals (CIs). Of the 4 SNPs that were studied, the hsa-mir-499 AG and GG genotypes were associated with a reduced risk of SCCHN (OR, 0.83; 95% CI, 0.69-0.99). When the 4 SNPs were combined according to putative risk genotype, the number of observed risk genotypes was associated with an increased risk of SCCHN in a dose-response manner with ORs of 1.0, 1.20, and 1.40 for individuals who had 0 or 1 risk genotypes, 2 or 3 risk genotypes, and 4 risk genotypes, respectively (P(trend) = .037). Specifically, the risk was 1.23-fold (95% CI, 0.98-fold to 1.56-fold) for individuals with 2 to 4 risk genotypes and 1.40-fold (95% CI, 1.02-fold to 1.92-fold) for individuals who had 4 risk genotypes compared with individuals who had 0 or 1 risk genotypes. This risk was more pronounced in men and in patients with oropharyngeal cancer. The combined risk genotypes of 4 common SNPs in pre-miRNAs were associated significantly with a moderately increased risk of SCCHN. Larger studies are needed to validate the current findings.

A novel, noninvasive, mRNA gene expression colon cancer screening methodology.

3631 Background: An mRNA gene expression methodology testing noninvasive rectal swab samples for a panel of 16 genes, representing 4 metabolic pathways, has shown promise for detecting colon and rectal cancers in pilot studies. To test the screening potential of this methodology, we conducted a case-controlled, single-center study using samples collected from patients with a negative screening colonoscopy (Controls; n = 71) and from patients with colorectal cancer (CRC; n = 38). Methods: Controls were defined as polyp- and cancer-free patients with no family history of CRC. Rectal swabs were collected, coded, preserved in RNAProtect Ò solution (Qiagen), and RNA was isolated using RNeasy mini kit (Qiagen). Real time RT-PCR using reverse transcriptase (Invitrogen) and random primers and Oligo (dT) was performed on samples pretreated with RNAase-free DNase. PCR amplification with specific oligonucleotide primers was performed using Applied Biosystems 7500 Fast Real-Time PCR Systems for 16 different markers including CXCR2, osteopontin, cyclo-oxygenase-1 and 2, proliferating peroxisome activating receptor alpha, delta and gamma, growth- related oncogene alpha and gamma, interleukin-8, mCSF, p21, c-myc, CD44, cyclin D and serum amyloid A1. Classification was based on using Mahalanobis distances with a threshold set to ensure at least 90% sensitivity. Results: For the 71 controls, the median age was 57 years (range 50-81) and 37 were women. For the 38 cancer patients, the median age was 67 years (range 50-89) and 19 were women. Adenocarcinomas were identified in the following colon locations: cecum (10), ascending (10), transverse (3), sigmoid (12), and rectosigmoid (3). AJCC tumor stage was I (9); II (15): III (13); and IV (1). The final analysis results in a test performance with 92% sensitivity and 68% specificity using mean M-Distance calculation. Sensitivity is 92% and specificity is 60% using maximum M-Distance calculation. Conclusions: This mRNA gene expression methodology would fulfill current guidelines for screening which require a minimum sensitivity and specificity of 50%. Future studies will aim to improve the specificity and validate the technology. Author Disclosure Employment or Leadership Position Consultant or Advisory Role Stock Ownership Honoraria Research Funding Expert Testimony Other Remuneration IntelliGene Scan IntelliGene Scan IntelliGene Scan IntelliGene Scan IntelliGene Scan

Pulmonary tuberculosis in patients with advanced head and neck cancer.

e16030 Background: Patients with head and neck cancer have more chances to suffer from pulmonary tuberculosis. Many patients with advanced head and neck cancer may have long-term survival after anticancer treatment. However, little has been mentioned about the consequence of tuberculosis in these patients. We addressed the impact of pulmonary tuberculosis on clinical outcome of patients with advanced head and neck cancer. Methods: From 2003 to 2007, in a tertiary referred medical center, we retrospectively reviewed the medical records of all 931 patients with active pulmonary tuberculosis, defined as a consistent chest imaging pattern and the growth of Mycobacterium tuberculosis in sputum or respiratory secretions. Totally 18 patients developed pulmonary tuberculosis within 2 years after the initial diagnosis of advanced head and neck cancer (stage III-IV). The demographic data, nutrition status, current cancer status and survival data were collected. Results: Seventeen of 18 patients were males. More than 90% of them have major risk factors (smoking, drinking, and betal-nut chewing) related to head and neck cancer. Eight patients with head and neck cancer were stage IV and all 18 patients received anticancer treatment. Eleven received combined modality therapy, 3 only radical operation, 4 operation alone. All patients received antituberculosis treatment. At the moment of anti-TB treatment, there were 6 patients with cancer-free status. Compared to these 6 patients, patients with uncontrolled cancer had a poor prognosis (survival time: 95% confident interval (CI) in cancer-free patients, 1358-2672 days; 95% CI in uncontrolled cancer: 55-482 days; p=0.0017 by log-rank test). In addition, serum albumin <3.0 g/dl also conferred to a worse prognosis (p=0.043). The cancer-free status and higher albumin levels were independent factors for a longer survival by multivariate analysis. Conclusions: Among patients with advanced head and neck cancer and active pulmonary tuberculosis, the status of uncontrolled cancer signifies a poor prognosis. Besides, poor albumin level in these patients may contribute to poor outcome. No significant financial relationships to disclose.

Combination of radiological and biochemical methods to assess bone mineral density of mandible in fully edentulous patients after chemotherapy: a 5-year prospective study

Background: Osteoporosis is of concern in breast cancer patients who are undergoing chemotherapy. This study compared the bone mineral density (BMD) index of the mandible and hip hinge between patients who were undergoing chemotherapy or breast cancer, and fully edentulous Chinese patients without cancer over a period of 5 years. Method: 120 fully edentulous patients with an average age of 69 who had undergone mastectomy for grade two or three non-metastatic invasive breast ductal carcinoma. This was followed by administration of 5-fluorouracil, cyclophosphamide, and doxorubicin. The 118 fully edentulous cancer-free patients were included as a control group. The first assessment point was 6 months after chemotherapy treatment. The BMD and panoramic and side views of the mandible were measured by γ-ray and the BMD of left hip hinge was measured using dual energy X-ray absorptiometry. The serum and urine level of bone-specific alkaline phosphatase (BAP), carboxyterminal cross-linked telopeptide of type I collagen (ICTP), as well as liver and renal function tests were determined. These examinations were performed annually for 5 years. Result: The cancer patients demonstrated a statistically significant (p < 0.05) increase in bone resorption of mandible and hip, and an increase in BAP and ICTP levels when compared with the control group. Although data were collected annually there was no statistical significance for the first 3 years. Conclusion: Breast cancer patients undergoing chemotherapy displayed significant resorption of mandibular bones compared with the healthy control, which might result in difficulties in fitting dentures, as it would cause pain and mucosal friction. Thus, concurrent therapy to decrease mandibular bone loss and special considerations in dentures are warranted for these patients.

Ferritin levels, inflammatory biomarkers, and mortality in peripheral arterial disease: A substudy of the Iron (Fe) and Atherosclerosis Study (FeAST) Trial

This study delineated correlations between ferritin, inflammatory biomarkers, and mortality in a cohort of 100 cancer-free patients with peripheral arterial disease (PAD) participating in the Veterans Affairs (VA) Cooperative Study #410, the Iron (Fe) and Atherosclerosis Study (FeAST). FeAST, a prospective, randomized, single-blind clinical trial, tested the hypothesis that reduction of iron stores using phlebotomy would influence clinical outcomes in 1227 PAD patients randomized to iron reduction or control groups. The effects of statin administration were also examined in the Sierra Nevada Health Care (SNHC) cohort by measuring serum ferritin levels at entry and during the 6-year study period. No difference was documented between treatment groups in all-cause mortality and secondary outcomes of death plus nonfatal myocardial infarction and stroke. Iron reduction in the main study caused a significant age-related improvement in cardiovascular disease outcomes, new cancer diagnoses, and cancer-specific death. Tumor necrosis factor (TNF)-alpha, TNF-alpha receptors 1 and 2, interleukin (IL)-2, IL-6, IL-10, and high-sensitivity C reactive protein (hs-CRP) were measured at entry and at 6-month intervals for 6 years. Average levels of ferritin and lipids at entry and at the end of the study were compared. The clinical course and ferritin levels of 23 participants who died during the study were reviewed. At entry, mean age of entry was 67 +/- 9 years for the SNHCS cohort, comparable to FeAST and clinical and laboratory parameters were equivalent in substudy participants randomized to iron reduction (n = 51) or control (n = 49). At baseline, 53 participants on statins had slightly lower mean entry-level ferritin values (114.06 ng/mL; 95% confidence interval [CI] 93.43-134.69) vs the 47 off statins (127.62 ng/mL; 95% CI, 103.21-152.02). Longitudinal analysis of follow-up data, after adjusting for the phlebotomy treatment effect, showed that statin use was associated with significantly lower ferritin levels (-29.78 ng/mL; Cohen effect size, -0.47 [t(df, 134) = 2.33, P = .02]). Mean follow-up average ferritin levels were higher in 23 participants who died (132.5 ng/mL; 95% CI, 79.36-185.66) vs 77 survivors (83.6 ng/mL; 95% CI, 70.34-96.90; Wilcoxon P = .05). Mean follow-up IL-6 levels were higher in dead participants (21.68 ng/mL; 95% CI, 13.71-29.66) vs survivors (12.61 ng/mL; 95% CI, 10.72-14.50; Wilcoxon P = .018). Ferritin levels correlated (Pearson) with average IL-6 levels (r = 0.1845; P = .002) and hsCRP levels (r = .1175; P = .04) during the study. These data demonstrate statistical correlations between levels of ferritin, inflammatory biomarkers, and mortality in this subset of patients with PAD.