bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4406: Clinical efficacious combined chemo/immunotherapy differently activates AKT pathway and functionality of gp100 and Melan-A specific T cell clones
Authors: Palermo Belinda , Donna Cosmo Di , Franzese Ornella , Bello Duilia Del , Gualtieri Novella , Imberti Luisa , Nuzzo Carmen , Proietti Enrico , Catricalà Caterina , Ferraresi Virginia , Nisticò Paola
Published: August 2012

A phase II randomized clinical study is ongoing in our Institution to prove the clinical efficacy of dacarbazine (DTIC) one day before peptide-vaccination (Melan-A and NY-ESO-1) in the prevention of melanoma relapse in clinically disease-free HLA-A2 patients. In a previous pilot study, this combination therapy (using Melan-A and gp100 peptides) increased the number of tumor-reactive long-lasting effector-memory CD8+ lymphocytes. To identify the mechanisms enhancing the immune response, induced by DTIC combined with peptide-vaccination, we analyzed the endogenous and treatment-induced antigen specific CD8 T-cell response at the clonal level. We analyzed the sequence of the TCR α-chain of these clones and the molecular results were correlated with the expression of CD27/CD28 co-stimulatory molecule, AKT activation and anti-tumor lytic activity. The combination of chemo/immunotherapy elicited in Melan-A-specific, but not in gp100 clones, a renewal of high-avidity/tumor-reactive T-cell clones, with a broadening TCR diversity in long-surviving patients, suggesting that the selection of immune-resistant tumor variants may be circumvented by this combination, and thus prevent tumor recurrence in melanoma. In gp100 clones, AKT activation (pSer473-AKT) canonically correlates with CD28 and/or CD27 expression, independent of the treatment while in Melan-A clones, lacking CD27 and CD28 expression, non-canonical AKT activation was only observed after the combination therapy. The identification of the extracellular stimuli and signaling pathway responsible for DTIC-mediated activation of the AKT signaling are currently under investigation.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4406. doi:1538-7445.AM2012-4406
Abstract 4409: A modular polymer platform for the delivery of cytokines is effective in the treatment of head and neck squamous cell carcinoma (HNSCC)
Authors: Lin Yuan , Suwarnasarn Arnold , Luo Jie , Elashoff David , Dubinett Steven , Wu Ben , John Maie St
Published: August 2012

Educational objective: At the conclusion of this presentation, the participants should be able to discuss the benefits of a modular polymer platform, which will improve the outcome for patients with advanced HNSCC. Objective: To evaluate the therapeutic efficacy of an immunomodulator secreting polymer platform in the treatment of HNSCC. Study Design: in vivo animal study. Setting: Academic research laboratory. Subjects and Methods: C3H/HeJ mice were injected with SCCVII/SF cells. After tumors reached 1 cm in size, animals then underwent surgery and were randomized to receive implantation of (1) no polymer; (2) plain polymer; (3) plain polymer with dendritic cells (DC) secreting the chemokine CCL21; (4) plain polymer with DC alone; and 5) plain polymer + daily CCL21 injection. Tumor size was measured until the mice were euthanized. At necropsy, the tumors were excised and weighed. Results: We were able to demonstrate that DC are viable and functionally secrete CCL21 up to 14 days in the polymer. DC-CCL21 secreting polymer effectively reduced tumor growth mice by over 16-fold (P < 0.01) as compared to control, plain polymer, and plain polymer + intratumoral CCL21 injection groups. Flow cytometry analysis revealed that there was a significant increase in CD4+ T cells, as well as a significant decrease in CD4+/CD25+ regulatory T cells in the tumor site, underlying the significant tumor regression observed. Conclusion: The major limitation for the clinical use of cytokines is the lack of an effective protocol for the sustained delivery of cytokines to the tumor milieu. We demonstrate the efficacy of a novel polymer platform in delivering DC-CCL21 to SCCA in a murine model. Our results indicate that this polymer may represent a new therapeutic modality for patients with HNSCC. Once this polymer platform is optimized we will plan for validation in the context of a prospective trial in patients with unresectable advanced or recurrent HNSCC.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4409. doi:1538-7445.AM2012-4409
Abstract 4412: Combining cytokine gene therapy with conventional cancer chemotherapyfor treatment of metastatic renal cell carcinoma
Authors: Yao Qian , Li Jianfeng , Liu Dexi
Published: August 2012

Renal cell carcinoma (RCC) accounts for approximately 3% of adult malignancies and 85-90% of kidney cancer. Treatment was generally aggressive involving nephrectomy and excision of the metastatic lesion. The current study was designed to evaluate a new strategy for treatment of RCC, using combination of cytokine gene therapy with conventional cancer chemotherapy. The study established tumor metastasis in Balb/c mice using Renca cells as a model1. Treatment was performed on tumor bearing mice with hydrodynamic gene delivery of plasmids containing either mIL-2 or mIL-12 gene, separately, or in combination with or without two weekly injection of 5-flououracil (5-FU) (20 mg/kg, ip). The results showed that the lung, liver and kidney were the primary sites of tumor growth. 5-FU is capable of inhibiting tumor growth in the liver and kidney and effective in slowing down tumor growth in the lung when treatment started early and with high frequency. mIL-2 gene therapy inhibited tumor growth in the liver and kidney. The effect became more obvious if treatment started early. However, IL-2 alone is unable to eliminate all liver tumors. mIL-2 gene therapy had almost no effect on tumor growth in lung. mIL-12 gene therapy significantly blocked cancer-induced weight loss. mIL-12 inhibited tumor growth in the liver and kidney, but IL-12 alone is unable to eliminate all liver and kidney tumors. In addition, IL-12 inhibited lung swelling induced by tumor, especially when treatment was started early. Cytokine gene therapy combined with chemotherapy is effective in blocking tumor-induced weight loss, and tumor growth in the liver, lung and kidney. These results provide direct evidence in support that combined therapy of cytokine gene therapy with traditional chemotherapy is effective in treating RCC. Key words: Renal cell carcinoma, gene therapy, IL-12, IL-2, 5- flououracil 1 Jianfeng Li, Qian Yao and Dexi Liu (2011) Hydrodynamic cell delivery for simultaneous establishment of tumor growth in mouse lung, liver and kidney. Cancer Biol Ther 12:737-741.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4412. doi:1538-7445.AM2012-4412
Abstract 4416: The pre-neoplastic lesion, mucin-depleted foci, reveals as de novo high-grade dysplasia in rat colon carcinogenesis
Authors: Cui Changxu , Takamatsu Reika , Doguchi Hiroshi , Matsuzaki Akiko , Saio Masanao , Yoshimi Naoki
Published: August 2012

Aberrant crypt foci (ACF) and mucin-depleted foci (MDF) have recently been recognized as the pre-neoplastic lesions in the colon of carcinogen-treated rodents. In the present study, we analyzed the sequential development of ACF and MDF histopathologically in the colon of rats from 5 to 40 weeks after DMH treatment. The number of ACF per colon increased over time during the experiment, and was much more than the numbers of tumor, while the number of MDF per colon remained unchanged from the early stage (the 5th week after carcinogen exposure), and approximate to that of tumor. The incidence of ACF, which was much higher than that of tumor, also increased gradually in a time-dependent way. The incidence of MDF, however, was similar to that of tumor and did not have significant change during the whole experiment. No the lesion as dysplasia with high-grade (DHG) or adenocarcinoma (AC) was found in any large ACF from the 5th to 40th week histopathologically, whereas all of the large MDF showed DHG or AC. Even at 5 weeks, MDF shows the appearance as DHG. We classified to two forms of MDF, a flat and a protruded MDF. At the 40 weeks, the number of a flat MDF per colon decreased significantly compared with that in the 20 weeks (p<0.05), but number of a protruded MDF per colon increased (p<0.01), and the percentage of DHG in a protruded MDF decreased but that of AC increased remarkably. In conclusion, MDF might develop into cancer through the so-called “de novo cancer” pathway.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4416. doi:1538-7445.AM2012-4416
Abstract 4419: The role of ABC transporters in PhIP-induced colon carcinogenesis
Authors: Durmus Selvi , van der Valk M.A. , Teunissen S.F. , Wagenaar Els , Beijnen Jos , Schinkel Alfred H.
Published: August 2012

ABC transporters such as BCRP, P-gp, Mrp2 and Mrp3 are thought to protect organisms from xenobiotics, including dietary carcinogens and anticancer drugs. These multidrug transporters reduce the oral uptake, systemic availability and tissue or cellular accumulation of transported substrates by mediating their extrusion from cells and the body in general. Therefore, they could have very important roles in the distribution of carcinogens to certain tissues and cell types, and carcinogen susceptibility. In this study, we aimed to investigate the role of ABC transporters in the tumorigenicity of a dietary carcinogen, PhIP (2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine), which is produced during high-temperature cooking of meat. We have performed a pharmacokinetic experiment in wild-type FVB and ABC transporter knockout strains (Bcrp;Mrp2;Mrp3-/- and Bcrp;Mdr1a/1b;Mrp2-/-) with 200 mg/kg orally administered PhIP and observed that ABC transporters have an impact on the distribution of PhIP metabolites to several organs. There was an accumulation of N-OH-PhIP, a potentially carcinogenic metabolite, in the liver of Bcrp;Mrp2;Mrp3-/- mice and a decrease in the levels of PhIP-5-sulfate, a surrogate marker for DNA exposure to reactive compound, in small intestine of Bcrp;Mrp2;Mrp3-/- and Bcrp;Mdr1a/1b;Mrp2-/- mice compared to wild-type FVB. For carcinogenesis experiments, we used a chemically-induced colon carcinogenesis model where wild-type FVB and ABC transporter compound knock-out mice received a single, oral administration of 200 mg/kg PhIP for initiation and seven days administration of Dextran Sodium Sulfate (DSS) in drinking water for promotion of the tumorigenesis process via colonic inflammation. During the whole experiment, mice were closely checked for their body weight loss, diarrhea and rectal bleeding. Our results showed that PhIP and DSS treatment together induced colonic tumors in all strains over 180 days. No tumor formation could be found in only PhIP-treated, only DSS-treated and negative control groups that received vehicle. Tumor incidence was lower in both knock-outs compared to the wild-type FVB when inflammation levels between the groups were corrected. Our results suggest that these ABC transporters are not protecting the intestinal tract from PhIP-induced carcinogenesis upon oral ingestion in mice, and perhaps even partially enhance PhIP-induced carcinogenesis.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4419. doi:1538-7445.AM2012-4419
Abstract 442: Increased miR-let7c level indicates transition from localized bladder cancer to metastasis
Authors: Meng Wei , Chakravarti Arnab , Lautenschlaeger Tim
Published: August 2012

Purpose/Objective(s): MicorRNAs are a group of short ribonucleic acid molecules with an average of 22 nucleotides and play an important role in regulation of gene expression. Because the microRNA profiles of tumor are related to cancer development and prognostication, it becomes a new frontier of cancer research. The current tools for studying microRNA like microarray have limited dynamic range and poor sensitivity, and Taqman microRNA assay has higher sensitivity but limited number of microRNAs. The SOLiD system provides a sensitive and robust assay for discovery and profiling of novel microRNAs using sequencing by ligation. The aim of this study is to find key miRNA which can provide insights into the cancer cell invasion and metastasis. Materials/Methods: 12 bladder cancer FFPE samples were used in this study, including 6 cases of stage N0, 3 cases of stage N1 and 3 cases of stage N2. Total RNA from FFPE blocks was isolated by Ambion RecoverAll Total Nucleic Acid Isolation kit according to manufacturer's instructions. The concentration and quality of total RNA were examined by Nanodrop 1000 spectrophotometer. The small RNA was enriched by flashPAGE fractionation and the microRNA library was built using Ambion SOLiD Small RNA Expression Kit with the barcode system (Life Technologies, CA, US). 35 bp long reads were generated on the SOLiD (version 3.0) sequencing system (Life Technologies) and were analyzed by using the SOLiD System Small RNA Analysis Tool (Life Technologies). Results: Analyzed samples were from male caucasian patients with a median age of 63.5 years. Tumors were all grade 3 urothelial carcinomas. Patients had T1-4 and M0-1 disease. We examined global miRNA expression in bladder cancer FFPE samples, and SOLiD results revealed that the expression of 10 microRNAs significantly differed between N0 stage and N1/2 stage. miR-let7c precursor and mature miR-let7c were upregulated in samples from patients with lymphnode metastasis. A previous study from Leite et al. showed that miR-let7c expression was similar in matched FFPE and frozen prostate cancer tissue with a Pearson's correlation coefficient of 0.927. Conclusion: miR-let7c expression is associated with the presence of lymphnode metastasis in high-grade urothelial bladder cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 442. doi:1538-7445.AM2012-442
Abstract 4421: PDE5 inhibitors suppress proliferation and augment the epithelial barrier by activating PKG2 in the colon
Authors: Wang Rui , Kwon In-Kiu , Subbaramia Sridhar , Browning Darren D.
Published: August 2012

Increasing cGMP levels has emerged as a potentially important suppressor of tumorigenesis in the colon, but the downstream signaling is poorly defined and a viable therapeutic strategy remains to be established. The present study has tested whether type 2 cGMP-dependent protein kinase (PKG2) mediates the homeostatic effects of cGMP in the colon. In addition, the feasibility of using phosphodiesterase 5 (PDE5) inhibitors therapeutically to increase cGMP levels in the colon mucosa has been examined. PKG2 is a central effector of cGMP in enterocytes, and consistent with a growth-inhibitory and differentiation-promoting role, knockout animals showed crypt hyperplasia and reduced mature goblet cells in the colonic mucosa. This phenotype is similar to guanylyl-cyclase C (GCC) knockout animals, which have also been reported to exhibit increased susceptibility to colon tumorigenesis. The possibility that PKG2 signaling might be activated pharmacologically, was examined using PDE5 inhibitors to regulate colonic proliferation and differentiation. These experiments demonstrated that intraperitoneal administration of clinically relevant PDE5 inhibitors provoked a rapid increase cGMP levels in the colon mucosa of mice. Moreover, prolonged treatment with PDE5 inhibitors led to reduced proliferation in the colonic crypts and a dramatic increase in mature goblet cells. Consistent with the importance of PKG2 in this process, the PDE5 inhibitors did not inhibit proliferation or increase goblet cell numbers in PKG2 knockout animals. The increase in goblet cell density in PDE5 inhibitor-treated animals was striking (50%, 85%, and 44% relative to controls in the ascending, transverse and descending colon respectively). Reduced basal epithelial apoptosis measured in PDE5 inhibitor treated animals suggested the the increased goblet cells could have a barrier-protective effect. To test this possibility further, the effect of PDE5 inhibitor treatment on epithelial damage by dextran sulfate sodium was measured. These studies demonstrated a significant attenuation of disease activity by PDE5 inhibitor treatment in this colitis model. In summary, our results demonstrate a central role for PKG2 in the regulation of proliferation and differentiation in the colon mucosa. In addition, this pathway can be exploited using PDE5 inhibitors, suggesting that these agents could be part of a novel therapeutic strategy for colon cancer prevention.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4421. doi:1538-7445.AM2012-4421
Abstract 4423: Transgenic liver expression of human arylamine N-acetyltransferase NAT2 restores sensitivity of male Nat1/2 null mice to 4-aminobiphenyl-induced liver tumorigenesis
Authors: Grant Denis M. , Brenneman Debbie , Sugamori Kim S.
Published: August 2012

We recently observed that male Nat1/2(-/-) mice, which are deficient in the arylamine N-acetyltransferases Nat1 and Nat2, have a significantly lower incidence and multiplicity of liver tumors than wild-type C57BL/6 mice after their neonatal exposure to the aromatic amine carcinogen 4-aminobiphenyl (ABP). Female mice showed low or no liver tumors from ABP exposure regardless of Nat1/2 status. The goal of the present study was to determine the effect of liver-selective transgenic expression of human NAT2 on ABP-induced liver tumorigenicity in male and female Nat1/2(-/-) mice. We generated a hNAT2tgNat1/2(-/-) strain of mice that express human NAT2 selectively in liver on a Nat1/2 null background, exposed male and female mice to either DMSO vehicle, a low dose (600 nmoles) or a high dose (1200 nmoles) of ABP during the second postnatal week, and assessed mice for tumor growth at one year of age. Male hNAT2tgNat1/2(-/-) mice had liver tumor incidences of 29% (low dose) and 61% (high dose), which more closely resembled those of C57BL/6 mice (60% and 69%, respectively) than those of Nat1/2(-/-) mice (0% and 35%, respectively). This suggests that human NAT2 is capable of substantially restoring the sensitivity of mice to ABP-induced liver tumorigenicity that is lost by the absence of mouse Nat1 and Nat2. On the other hand, female hNAT2tgNat1/2(-/-) mice were still as resistant to liver tumors as C57BL/6 or Nat1/2(-/-) mice. No tumors were observed in any other organs. These results suggest that the human and mouse NAT enzymes play similar roles in enhancing liver tumor growth, and that the mechanism of female protection against ABP-induced tumor growth is unrelated to NAT status. However, neither the sex nor the NAT status of the mice appear to be correlated with the ability of ABP to produce acute DNA-damaging or mutation-inducing effects in liver. This suggests that the role of the human and mouse NAT enzymes in promoting liver tumor growth in mice may be due to a mechanism that is independent of their ability to influence the process of ABP metabolic activation and subsequent DNA damage leading to tumor initiation.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4423. doi:1538-7445.AM2012-4423
Abstract 4473: Urinary biomarkers of exposure to 1,3-Butadiene and its bioactivation to DNA-reactive metabolites
Authors: Kotapati Srikanth , Sangaraju Dewakar , Walker Vernon E. , Swenberg James A. , Tretyakova Natalia Y.
Published: August 2012

1,3-butadiene (BD) is one of the most potent and abundant carcinogens present in cigarette smoke. BD is metabolically activated to 3,4-epoxy-1-butene (EB), 1,2,3,4-diepoxy butane (DEB), hydroxymethylvinylketone (HMVK), and 3,4-epoxy-1,2-butanediol (EBD), which can react with DNA nucleobases and proteins to form promutagenic adducts. Alternatively, these electrophilic metabolites can be detoxified via hydrolysis or conjugation with glutathione. The glutathione conjugates of these metabolites are ultimately excreted in urine as 1-hydroxy 2-(N-acetylcysteinyl)-3-butene (MHBMA), 1,4-bis-(N-acetylcysteinyl)butane-2,3-diol (bis-BDMA), 1,2-dihydroxy-4-(N-acetylcysteinyl)-butane (DHBMA), and 1,2,3-trihydroxy-4-(N-acetylcysteinyl)-butane (THBMA), respectively. Since epoxide metabolites of BD are thought to be responsible for its adverse health effects, it is important to develop specific biomarkers of their formation in vivo to be used in molecular epidemiology studies. While MHBMA, DHBMA, THBMA have been previously detected in smoker's urine, bis-BDMA has not been reported. In the present study, novel stable isotope dilution HPLC-ESI-MS/MS methods for accurate and precise quantification of MHBMA, DHBMA, THBMA, and bis-BDMA in urine have been developed. Dose dependent formation of all BD-mercapturic acids including bis-BDMA was observed in urine samples from male and female F344 rats exposed to 0, 62.5 and 200 ppm of BD for 2 weeks by inhalation. The relative amounts of MHBMA, DHBMA, THBMA, and bis-BDMA in rat urine were 20:50:50:1. Furthermore, MHBMA, DHBMA, and THBMA were detected in urine samples from known smokers and non-smokers, with significantly higher concentrations observed in smokers. The mean levels of these BD-mercapturic acids in smokers were 3, 340, and 27 ng/mg creatinine, respectively. In contrast, bis-BDMA could not be detected in human urine samples, probably a result of the inefficient DEB formation in humans. These methods are now being applied to a multi-ethnic cohort study to identify any differences in metabolism that contribute to ethnic and inter-individual differences in lung cancer incidence.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4473. doi:1538-7445.AM2012-4473
Abstract 4474: Occupational exposure to benzene and leukocyte telomere length
Authors: Bassig Bryan , Zhang Luoping , Cawthon Richard , Yin Songnian , Li Guilan , Rappaport Stephen , Hu Wei , Smith Martyn T. , Rothman Nathaniel , Vermeulen Roel , Lan Qing
Published: August 2012

Benzene is an important industrial solvent and a ubiquitous environmental contaminant. IARC has classified benzene as a human leukemogen (Group 1) and a possible lymphomagen. We have previously demonstrated that exposure to benzene results in hematotoxic effects in a cross-sectional study, and that longer telomere length may be associated with non-Hodgkin lymphoma (NHL) in a prospective study conducted in the general population. To further explore the biologic plausibility that benzene may be a lymphomagen, we tested if leukocyte telomere length was altered in a cross-sectional biomarker study of workers occupationally exposed to benzene in Shanghai, China. Forty-four benzene-exposed workers were enrolled from factories that used benzene, along with 44 age and gender-matched controls from two workplaces in the same geographic area that did not use benzene or other chemicals associated with bone marrow toxicity. Personal exposure measurements were obtained for each exposed worker using passive 3M organic vapor monitoring badges. Blood samples were collected from each worker, and leukocyte telomere length was measured using a quantitative PCR procedure. Linear regression using the natural logarithm of telomere length and adjusting for potential confounders was used to test for differences in telomere length between exposed workers and controls. The median benzene personal air level was 31 ppm in the exposed workers. Compared to unexposed workers, the mean leukocyte telomere length was increased by about 8% in all benzene-exposed workers (p=0.05). There was a 4% increase in workers exposed to < 31 ppm and a 12% increase in workers exposed to > 31 ppm and a statistically significant dose-dependent increase in telomere length with increasing levels of benzene exposure in air (ptrend = 0.01). Some preliminary evidence has suggested that benzene exposure may stimulate telomerase expression in lymphocytes and other cell lines. Given that longer telomere length has been associated with risk of NHL, our findings provide some support for the biologic plausibility that benzene is a lymphomagen but require replication in larger studies.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4474. doi:1538-7445.AM2012-4474
Abstract 4477: Progesterone (P4) and testosterone (T) concentrations in random fine needle aspirates of the breast in pre- and postmenopausal women
Authors: Heinz Richard E. , Kenney Kara , Lee Oukseub , Iyer Sivaraman , Khan Seema A. , Chatterton Robert T.
Published: August 2012

Background: The availability of steroid hormones in the breast may be an important measure of the potential for proliferation of hormone-dependent breast cancer, and because retention of hormones may be influenced by serum levels, in situ formation, and the presence of binding proteins and lipids in the tissue, measurements of tissue concentrations are more informative than serum concentrations alone. Methods: Subjects were normal women, aged 36-65 yr, without breast cancer and not taking hormones. Tissue was obtained by fine needle aspiration of the breast. The cellular pellet was saved for analysis of DNA, and lipids were extracted from the supernatant including the fatty layer. Previous work had shown the cellular pellet contained a very little of the total steroids of the sample. Triglycerides were precipitated from 90% methanol at –20 °C and the remaining lipids were applied to a C18 HPLC column for purification in a gradient of methanol and acetonitrile in phosphate buffer. Fractions containing P4 and T were analyzed by immunoassays. The total DNA in the cellular pellet was isolated and quantified by UV absorbance. The steroid content was expressed as either per mg of purified lipid or per microgram of DNA. The data, when expressed as the natural log of the concentrations, followed a normal distribution. Differences in pre- and postmenopausal concentrations were analyzed by group t-tests. Results: The geometric means of the hormone concentrations are shown in the table. Comment: Despite the almost complete loss of progesterone in serum of postmenopausal women, the concentrations in the tissue are reduced but remain, on average, at more than 25% of the premenopausal level, somewhat more than the previously reported percentage in NAF of 5%. Relationships within individual women to other risk factors will be explored. Testosterone is also maintained at a relatively high level in the tissue of postmenopausal women, and may serve as a substrate for aromatase.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4477. doi:1538-7445.AM2012-4477
Abstract 4486: Mitochondrial DNA copy number is associated with colorectal cancer risk
Authors: Thyagarajan Bharat , Wang Renwei , Barcelo Helene , Koh Woon-Puay , Yuan Jian-Min
Published: August 2012

Background: Mitochondrial DNA (mtDNA) copy number variation is common in colorectal tumors and one case control study showed mtDNA copy number to be associated with increased colorectal cancer risk. Data on mtDNA copy number and colorectal cancer risk in prospective cohorts is lacking. Objectives: We evaluated the association between mtDNA copy number in peripheral blood and colorectal cancer risk among participants of the Singapore Chinese Health Study. Methods: We conducted a nested case-cohort study of 422 colorectal cancer cases (168 incident cases with pre-diagnostic blood and 254 prevalent cases with post-diagnostic blood) and 880 controls who were free of colorectal cancer at blood draw. The relative mtDNA copy number was measured using real time PCR. Analysis of covariance and logistic regression methods were employed to examine the association between mtDNA copy number and colorectal cancer risk. Results: mtDNA copy number was significantly higher among colorectal cancer cases than controls (geometric mean: 0.62 vs. 0.50, p<0.0001). There was a U shaped relationship between the mtDNA copy number and colorectal cancer risk. Individuals in the low and high mtDNA copy numbers were at increased risk of colorectal cancer; odds ratios (95% confidence interval) for developing colorectal cancer for the lowest and highest quartile of mtDNA copy numbers were 1.83 (1.16-2.90) and 3.54 (2.25-5.55), respectively, compared to those in the second quartile of mtDNA after adjustment for potential confounders (p for curvilinear trend <0.001). The lowest mtDNA copy number quartile was associated with colorectal cancer only in the pre-diagnostic blood samples (OR=3.22, 95% CI:1.51-6.85) but not in the post-diagnostic blood samples (OR=1.36, 95% CI:0.80-2.32), implying that cancer status impacts the mtDNA copy number-colorectal cancer risk association. Conclusions: This is the first study to show a prospective association between mtDNA copy number and colorectal cancer risk. Further studies are needed to confirm this association and understand the biological significance of these observations.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4486. doi:1538-7445.AM2012-4486
Abstract 4491: Transforming growth factor (TGF) ≤ pathway and clinical outcome of pancreatic cancer
Authors: Javle Milind M. , Dong Xiaoqun , Tan Dongfeng , Li Yanan , Kar Siddhartha P. , Baladandayuthapani Veera , Weatherly Jacqueline , Krishnan Sunil , J Tzu-chuan , Huang  , Fogelman David R. , Abbruzzese James L. , Wolff Robert A. , Li Donghui
Published: August 2012

Background: TGFβ acts both as a tumor suppressor or tumor promoter depending upon cellular context and stage of tumor progression. TGFβ exerts its effects through the TGFβR1/ R2 receptors and the SMAD transcription regulators. We investigated the prognostic value of TGFβ signaling biomarkers in pancreatic cancer. Methods: We measured plasma TGFβ1 level using Meso Scale Discovery Multi-array® Human TGFβ1 Assay in 643 pancreatic cancer patients. TGFβR2 (Novus) and SMAD4 (Proteintech) protein expression were measured in 86 biopsies using immunohistochemisty (IHC). IHC intensity was scored 0-3. Percentage IHC-positive cancer cells were 0-3 (0: 0%+, 1: 1-10%+, 2: 10-25%+, 3: >25%+). Final IHC score was measured as intensity × percentage. We also genotyped 3 SNPs of TGFB1 gene using the Taqman assay. Kaplan-Meier and log-rank tests were used to analyze overall survival (OS) by TGFβ1 level. Multivariate Cox proportional hazards models with relevant clinical covariates were used to assess the relationship between IHC score and OS. Results: In patients with locally advanced and metastatic disease (n = 355), there was a significant association of plasma TGFβ1 with OS. OS of the top quartile of TGFβ1 levels (>19.05 ng/mL) vs. those with low level was 27.7 weeks vs. 40 weeks, respectively [log-rank p = 0.0125, adjusted for baseline CA 19-9 and performance status (PS)]. No significant association between plasma TGFβ1 and OS was noted in surgical patients (n = 288). In the multivariate Cox model, after adjusting for baseline stage, CA 19-9, PS, age and TGFβR2 expression, complete loss of SMAD4 expression (IHC score 0) was significantly associated with lower OS [HR (95% CI): 1.85 (1.06-3.23), p = 0.03]. Progressive disease on gemcitabine-based therapy was more likely in SMAD4 IHC 0 group as compared with higher score (46.5% vs. 38.1% progressed; chi square p = 0.069). TGFB1 –1346T>C CT/TT genotype correlated with shorter OS [HR (95% CI): 1.21 (1.02-1.45), p = 0.03] after adjusting for clinical variables. Conclusions: This large retrospective study suggests an important stage-dependant role of the TGFβ pathway in pancreatic cancer. Targeting this pathway in advanced pancreatic cancer may have therapeutic relevance.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4491. doi:1538-7445.AM2012-4491
Abstract 4493: Serum X-linked inhibitor of apoptosis protein (XIAP) predictive recurrence of renal cell carcinoma
Authors: Hongo Fumiya , Takaha Natsuki , Kimura Yasunori , Nakamura Terukazu , Mikami kazuya , Miki Tsuneharu
Published: August 2012

Purpose: The X-linked Inhibitor of Apoptosis (XIAP) is associated with cell survival by blocking caspase-mediated apoptosis. The expression and prognostic significance of XIAP in renal cell cancer (RCC) has been rarely studied. To our knowledge, no report on the serum XIAP levels from RCC patients has been published. In this study, we examined serum XIAP levels of RCC patients and normal individuals and evaluated its utility as biomarker. Methods: Peripheral blood samples were obtained from 84 patients (59 men and 25 women, median age; 63 years [36-85]) with RCC before surgery. All the patients underwent radical or partial nephrectomy. Blood samples were also collected from 52 healthy controls. The serum TRAIL levels were measured by a sandwich enzyme-linked immunosorbent assay (Gencompare, TiterZyme®). Cut off value was calculated by ROC analysis. Results: The serum XIAP levels in patients with RCC were higher than those of normal control individuals (328.3 pg/ml vs 156.2 pg/ml, P<0.001). The progression free survival rate of the RCC patients with high serum XIAP was shown to be significantly lower than that of those with low serum XIAP in Grade 1-2 group (P < 0.05). Conclusions: Serum XIAP level is higher in renal cell cancers compared to normal control. High XIAP expression is not directly associated with tumor stage. These results suggest that it may be used as a novel prognosticator and a potential target for renal cell cancer diagnosis and therapy.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4493. doi:1538-7445.AM2012-4493
Abstract 4498: Prognostic significance of nuclear factor kappa B expression in human papillomavirus-associated oropharyngeal cancer
Authors: Zhang Hongzheng , Hu Zhongliang , Phelan Vanessa , Liu Yuan , Müller Susan , Saba Nabil F. , Beitler Jonathan J. , Khuri Fadlo R. , Chen Georgia Z. , Shin Dong M.
Published: August 2012

Nuclear factor kappa B (NFκB) has been implicated in the development of head and neck squamous cell carcinoma (HNSCC) from premalignant changes to tumor progression and recurrence. However, the prognostic significance of cytoplasmic or nuclear localization of NFκB expression in human papillomavirus (HPV)-associated HNSCC has not been investigated largely due to limited patient samples. We hypothesized that cytoplasmic and/or nuclear expression of NFκB may be a poor prognostic factor for tumor recurrence and overall survival (OS) regardless of p16 status. We conducted a retrospective study with archival formalin-fixed paraffin-embedded tumor biopsies collected from 188 patients with oropharyngeal SCC between October 1994 and February 2008 from Emory University Department of Pathology. Median follow-up was 21.7 and 58.0 months for p16- and p16+ patients, correspondingly. Tumors were evaluated for p16 expression (a surrogate marker for HPV) by immunohistochemistry (IHC). Cytoplasmic expression of NFκBp65 by IHC or total expression combining cytoplasmic and nuclear localization were significantly greater in p16- patients (N=58) than p16+ patients (N=130), p<0.05. High cytoplasmic expression of NFκBp65 was significantly associated with poor OS (p=0.0276) and the total expression was associated with recurrence (p=0.0081) by log rank test, whereas nuclear NFκBp65 displayed weak association. Positive nuclear expression significantly correlated with smoking history (p=0.025). In conclusion, p16+ and low total NFκBp65 expression is associated with the best while p16- and high total NFκBp65 is associated with the worst OS in this patient cohort (Table I). (The study was supported by Head and Neck SPORE Grant P50CA128613).

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4498. doi:1538-7445.AM2012-4498
Abstract 45: The clinical significance of CD163 in patients with hepatocellular carcinoma who underwent curative resection
Authors: Hui-Chuan Sun , Kong Ling-Qun , Zhu Xiao-Dong , Chai Zong-Tao , Tang Zhao-You
Published: August 2012

Background We have found that the abundance of peritumoral CD68+ macrophages was associated with patients’ prognosis after curative resection of hepatocellular carcinoma (HCC). However, the role of the pro-tumoral or tumoricidal macrophages is still not clear. Methods In this study, the clinical significance of immunostaining of CD163, a marker of alternatively activated macrophages (M2), in tumor and peritumoral liver tissue from a cohort of 295 Chinese HCC patients who underwent curative resection was evaluated and compared with that of the immunostaining of CD68. The clinical significance of soluble form of CD163 (sCD163) in peripheral blood was also evaluated in another cohort of 107 HCC patients. Results The count of CD163+ cells was well correlated with and much higher than that of CD68+ cells in both tumors and peritumoral liver tissues respectively, which was consistent with the observation from immunostaining by CD68 or CD163 antibody on the consecutive sections. Patients with a higher count of peritumoral CD68+ cells was associated with poor recurrence-free survival and overall survival (P = 0.004 and, P = 0.001, respectively), whereas the counts of tumor and peritumoral CD163+ cells did not. Plasma sCD163 level was associated with active hepatitis-related factors (e.g. serum alanine aminotransferase, serum aspartate aminotransferase, γ-glutamyl transpeptidase and serum alkaline phosphatase). Conclusions CD163+ cell count is of limited significance to predict patients’ prognosis in HCC. Plasma sCD163 was more likely a factor related with active hepatitis rather than tumor related features.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 45. doi:1538-7445.AM2012-45
Abstract 4500: Clinical significance of survivin expression in urothelial carcinoma of the bladder
Authors: Wang Yuan-Hung , Hsieh Hsiao-Yen , Hsu Cheng-Da , Tung Chun-Liang , Shen Kun-Hung , Shen Cheng-Hung
Published: August 2012

Objective: Survivin is one of the inhibitors of apoptosis protein family and plays an important role in carcinogenesis. Survivin also has been reported to be related with poor clinical outcome in various malignancies. The aim of this study is to examine the association between survivin expression and clinical outcome in urothelial carcinoma of the bladder. Materials and methods: A total of 56 histopathologically confirmed urothelial carcinoma of bladder (UCB) patients were recruited from the Department of Urology of Chiayi Christian Hospital and Chi-Mei Medical Center from August 2006 to May 2007. Immunohistochemistry (IHC) analysis was performed to determine the expression level of survivin in tumor and adjacent normal tissues. One normal human urothelial cell: HUC4449 and nine urothelial carcinoma cell lines which are TSGH8301, 5637, BFTC905, BFTC909, HT1376, TCC SUP, J82, T24 and HT1197 for survivin mRNA analysis by quantitative real-time PCR. The –31 C/G polymorphism in survivin gene was determined using PCR-RFLP method. The effects of a higher expression level of survivin on disease-free survival (DFS) and overall survival (OS) were estimated using Cox proportional hazard models. Results: The frequency of low, moderate and higher expression of survivin in tumor tissues of UCB were 16.1%, 39.3% and 44.6%, respectively. The frequency of high expression of survivin was significantly higher in muscle-invasive tumors (66.6%) than in non-muscle invasive tumors (34.2%) (P=0.042). The frequency of high expression of survivin was significantly higher in poor-differentiated (85.7%) than in moderate- differentiated tumors (30.8%) (P= 0.0014). The survivin mRNA expression (Log2 fold change were 3.41∼5.30) in various grade (G2-G4) urothelium carcinoma cell lines were all higher than normal human urothelial cell. Moreover, the higher frequency of risk genotypes (C/C+C/G) was observed in median and high survivn expression groups. In a multivariate analysis, high survivin expression was predictive of inferior OS (HR= 12.3, P=0.02). Conclusion: These findings suggest that the high expression of survivin is not only associated with tumor stage and grade of tumors but also may be predictive of inferior overall survival of urothelail carcinoma of the bladder. Keywords: survivin; bladder; urothelial carcinoma; immunohistochemistry, disease-free survival; overall survival

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4500. doi:1538-7445.AM2012-4500
Abstract 4501: Energetic risk and prostate cancer-specific and all-cause mortality in two large cohorts of men with localized prostate cancer
Authors: Ma Jing , Giovannucci Edward , Nguyen Paul L. , Qiu Weiliang , Chavarro Jorge , Rosner Bernard , Cook Nancy R. , Platz Elizabeth A. , Gaziano J. Michael , Pollak Michael , Stampfer Meir J.
Published: August 2012

Objective: To examine whether high energetic risk predisposes men with localized prostate cancer (PC) to higher risk of PC-specific mortality or all-cause mortality. Methods: Among men with localized PC, prediagnostic body mass index (BMI) and plasma C-peptide (a marker of insulin production) were available for 691 men (301 overall deaths, 78 PC deaths) in the Physicians’ Health Study (discovery set) and 1111 men (262 overall deaths, 56 PC deaths) in the Health Professionals Follow-up Study (validation set). High energetic risk was defined as BMI 25-29.9 kg/m2 and C-peptide in the highest quartile or BMIα30 kg/m2. We used the Cox-regression model to estimate risk, adjusting for age, smoking, diabetes, and D'Amico risk defined by stage, Gleason grade and prostate-specific antigen (PSA). Results: Results were similar in both cohorts; the combined hazard ratio (HR)s (95% confidence interval, CI) associated with high energetic risk (25% of study population) were 2.2 (1.5-3.3) for PC-specific mortality and 1.4 (1.1-1.7) for all-cause mortality. The risk of PC-specific mortality according to low- intermediate-, or high-D'Amico risk were 1.0 (reference), 2.6, 6.4 for low energetic risk, and 5.0, 6.1, 9.4 for high energetic risk. The corresponding HRs for all-cause mortality were 1.0 (reference), 1.3, 2.1, 2.1, 2.1, and 2.3. Compared to D'Amico risk alone, incorporating energetic risk significantly improved the predictability of PC-specific mortality (C-statistic from 0.72 to 0.78, P<0.001). Conclusion: Adding energetic risk to D'Amico risk groups significantly improved prediction of PC-specific and all-cause mortality, and identifies 20% D'Amico “low-risk” patients who may be poor candidates for active surveillance.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4501. doi:1538-7445.AM2012-4501
Abstract 4502: DNA ploidy in tumor correlates with age, and explains some of the poor prognostic impact of age in endometrial carcinomas
Authors: Mauland Karen K. , Wik Elisabeth , Werner Henrica MJ , Trovik Jone , Salvesen Helga B.
Published: August 2012

Background: Endometrial cancer incidence is increasing in industrialized countries, and the demand for better prognostic markers in order to individualize treatment is emerging. DNA ploidy has previously shown independent prognostic impact in smaller studies. In this study, we wanted to investigate the prognostic impact of ploidy in primary endometrial tumors, and the link between DNA ploidy and patient age at diagnosis in relation to traditional clinicopathologic prognostic variables. Study design: 663 patients treated for endometrial carcinoma at Haukeland University Hospital, Norway, from 1981 through 2010 were included. DNA ploidy in fresh tumor specimens was measured by flow-cytometry. Comprehensive clinical and histopathologic data, treatment and complete follow-up were collected. The software PASWStatistics18 was used for statistical analysis. Results: DNA aneuploidy was significantly correlated to high patient age at diagnosis (p<0.001). It was also correlated to non-endometrioid histology and high-grade tumors (p<0.001) and high FIGO stage (p=0.012). In univariate survival analysis (Kaplan Meier), the 5-year disease specific survival of patients with diploid tumors was 89%, versus 65% for aneuploid tumors (p<0.001). In Cox regression analysis, aneuploidy was significantly associated with poor survival (HR=2.4, 95%CI 1.6 - 3.8, p<0.001), adjusted for FIGO stage, age, grade, and histologic subtype. Interestingly, the prognostic impact of patient age diminished when ploidy status was included in the Cox analysis. Conclusion: DNA ploidy estimation in endometrial carcinoma tumor tissue is highly associated with patient age at diagnosis, as well as prognosis. The prognostic impact of age decreases when adjusting for ploidy status in tumor.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4502. doi:1538-7445.AM2012-4502