bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 3707: Sulindac sulfide inhibits growth and induces apoptosis of human colon tumor cells by a cGMP-dependent pathway leading to suppression of β-catenin transcription activity
Authors: Li Nan , Tinsley Heather , Whitt Jason , Gary Bernard , Lu Wenyan , Li Yonghe , Keeton Adam , Piazza Gary
Published: June 2012

Sulindac and other nonsteroidal anti-inflammatory drugs have shown promise for chemoprevention of colorectal cancer, but toxicities arising from cyclooxygenase (COX) inhibition limit their clinical use. Previous reports suggest that sulindac may exert its antitumor effects by a COX-independent target involving cyclic guanosine monophosphate (cGMP) phosphodiesterase (PDE) inhibition, although the specific isozymes involved and mechanism have not been well defined. In this study, we show that sulindac sulfide (SS) inhibits the growth of human colon tumor cells, HCT116, HT29 and SW480 with IC50 values of 60-80μmol/L. Within this concentration range, SS did not affect the growth of human-derived normal colonic epithelial cells, NCM460. Furthermore, SS induced apoptosis of colon tumor cells but not NCM460 cells. SS inhibited cGMP PDE activity in colon tumor cell lysates with minimal effect on cGMP PDE activity in NCM460 cell lysates. By determining the sensitivity of all eleven PDE isozymes to SS, the cGMP-specific PDE5 was found to be the most sensitive with an IC50 value for enzyme inhibition of 38µmol/L. In addition, PDE5 was found to be overexpressed in colon tumor cell lines compared with NCM460 cells. Knockdown of PDE5 expression in colon tumor cells with siRNA resulted in a significant reduction of tumor cell growth and increased apoptosis. Moreover, SS increased intracellular cGMP levels, activated protein kinase G (PKG) and suppressed the expression of β-catenin in colon tumor cells. SS also inhibited the transcriptional activity of β-catenin and reduced the expression of the β-catenin regulated proteins cyclin D1 and survivin, which are important mediators of tumor cell proliferation and apoptosis, respectively. Our results suggest that the chemopreventive properties of sulindac are closely associated with PDE5 inhibition and involve the activation of cGMP signaling to suppress β-catenin mediated transcriptional activity. Funding provided by NIH/NCI grants CA131378 and CA148817.

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 3707. doi:10.1158/1538-7445.AM2011-3707
Abstract 3709: Molecular targets of total methanolic and ethyl acetate fraction of triphala in colon cancer
Authors: Ravoori Srivani , Higashi Richard M. , Gupta Ramesh C. , Vadhanam Manicka V.
Published: June 2012

We previously demonstrated that the methanolic extract of triphala, a herbal preparation of a mixture of three fruits, namely, Terminalia chebula, Emblica officinalis and Terminalia bellerica was effective against colon cancer cells. In order to identify its mechanism of action, HCT-116 cells (1×106 cells) were seeded in 25mm2 petri dishes and treated with varying concentration of total extract. Cells stained using Alexa Fluor 488 Annexin V/Dead Cell Apoptosis assay revealed a dose dependent increase in early apoptosis which plateaued with 200 µg/ml, and a dose dependent increase in late apoptosis upto 400 µg/ml. This and previous finding of down-regulation of cyclin D1 suggested that the inhibitory action of triphala is facilitated both by antiproliferation and induction of apoptosis. In addition, significant downregulation of select upstream and downstream molecules of NFκB signaling was observed. In order to identify the bioactive components of the mixture, the total methanolic extract was subfractionated with various organic solvents and tested for antiproliferative activity using HCT-116 cells. The efficacy was determined to be in the following order: ethyl acetate fraction>total methonolic fraction>chloroform fraction>butanol fraction>hexane fraction>residual aqueous fraction. When normal colon cells CRL-1459 (5×103 cells) were treated with either the total methanolic or ethyl acetate fraction, the IC50 was found to be 7 fold higher for both the fractions compared to colon cancer cells, suggesting that the antiproliferative activity is specific to malignant cells. FTICR-MS analysis of the effective fractions revealed enrichment of most of the phenolics such as gallic acid, ellagic acid, and several other tannins into the ethyl acetate fraction. In order to identify the molecular targets of the ethyl acetate fraction, HCT-116 (1×106 cells) were seeded in 6 well plates and treated with LPS, 100 or 200 µg/ml ethyl acetate extract of triphala or both for 6 hours. qRT-PCR analyses revealed that the ethyl acetate fraction is more effective in down-regulating both constitutive and induced expression of TNFα and COX2 compared to the total methanolic extract. In addition, down-regulation of LPS-induced NFκB expression was more pronounced with ethyl acetate extract compared to methanolic extract. Ethyl acetate extract was also effective in downregulating the cell cycle regulatory molecule cyclin D1. However, the total extract was more effective in modulating the expression of IL-6 and IFNγ, suggesting that the various fractions of triphala may have distinct targets of colon carcinogenesis.

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 3709. doi:10.1158/1538-7445.AM2011-3709
Abstract 371: Comparison of toxicity and study design issues of nanoparticle and small molecule anticancer agents in preclinical models and phase I clinical trials
Authors: Morgan Katherine P. , Caron Whitney P. , Walsh Mark D. , Zamboni Beth A. , Zamboni William C.
Published: June 2012

Background: Carrier-mediated agents (CMA) are classified as nanoparticles, nanosomes (nanoparticle sized liposomes) and conjugates. Anticancer CMA offer many unique advantages over their traditional small molecule (SM) counterparts, including improved solubility, longer duration of exposure, tumor-selective delivery, increased antitumor response and reduced toxicity. The interpatient variability in pharmacokinetics (PK) and pharmacodynamics (PD) associated with nanoparticles are greater than that observed with SM. The starting dose for phase I studies of SM and CMA anticancer agents is based on the toxicity profile of the most sensitive species (e.g. rat or canine). However, the optimal animal model for toxicologic and pharmacologic studies of CMA is unclear. Our preliminary data suggests that dogs are highly sensitive to nanoparticles and thus may not be an appropriate animal model in determining the starting dose of CMA for phase I clinical trials. We evaluated preclinical toxicology data and how this affected the design and progression of phase I studies of CMA compared to SM anticancer agents.

Methods: In preclinical studies, the maximum tolerated dose (MTD) in rats and dogs was evaluated for CMA and their respective SM. In phase I clinical trials of CMA and their respective SM in patients with advanced solid tumors, the basis for starting dose, the number of dose escalations, number of patients and the ratio of MTD to starting dose was determined.

Results: Starting dose in phase I studies of CMA was based on dogs.

Conclusions: The degree of dose escalation from starting dose to MTD was significantly greater for CMA compared to SM drugs. This was also associated with a significantly greater number of dose levels, patients, and time required to complete phase I studies of CMA compared to SM. These findings necessitate the need to identify the most appropriate preclinical animal model to use to evaluate CMA toxicity in phase I studies.

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 371. doi:10.1158/1538-7445.AM2011-371
Abstract 3718: Well-done meat intake and meat-derived mutagen exposures in relation to breast cancer risk: The Nashville Breast Health Study
Authors: Fu Zhenming , Deming Sandra L. , Fair Alecia M. , Shrubsole Martha J. , Wujcik Debra M. , Shu Xiao-ou , Kelley Mark C. , Wei Zheng
Published: June 2012

Previous studies of the association of meat intake and meat-derived mutagen exposure with breast cancer risk have produced inconsistent results. We evaluated this association in a population-based case-control study of incident breast cancer conducted in Nashville,TN, United States, including 2,386 breast cancer cases and 1,703 healthy women controls. Telephone interviews were conducted to obtain information related to meat intake including amount, cooking methods, and doneness levels, as well as other known or hypothesized risk factors for breast cancer. Unconditional logistic regression was used to derive odds ratios (ORs) after adjusting for potential confounders. High intake of red meat was associated with a significantly elevated risk of breast cancer (P-trend <0.001). The association was particularly strong for high intake of well-done red meat (P-trend <0.001), with an adjusted OR of 1.5 (95% CI = 1.3-1.9) for the highest versus the lowest quartile. Associations between red meat and breast cancer risk were stronger for postmenopausal women than for premenopausal women. Meat-derived mutagens such as 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx) and 2-amino-3,4,8-trimethylimidazo[4,5-f]quinoxaline (DiMeIQx), were significantly associated with increased breast cancer risk among postmenopausal women only (P-trend = 0.002 and 0.003, respectively). The results from this study provide strong support for the hypotheses that high red meat intake and meat-derived mutagen exposure may be associated with an increase in breast cancer risk.

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 3718. doi:10.1158/1538-7445.AM2011-3718
Abstract 3724: Use of aspirin, other nonsteroidal anti-inflammatory drugs (NSAIDs), acetaminophen and risk of postmenopausal breast cancer
Authors: Zhang Xuehong , Smith-Warner Stephanie , Giovannucci Edward , Rosner Bernard , Willett Walter , Hankinson Susan
Published: June 2012

Background Aspirin is widely used for cardiovascular disease prevention and has been consistently shown to be associated with a 20-30% lower risk of colon cancer. However, the associations between uses of aspirin, other NSAIDs, and acetaminophen and breast cancer incidence are uncertain. In addition, limited epidemiological studies have assessed these associations with breast cancer subtypes defined by hormone receptor status. Further, no known prospective cohort studies have been conducted on other clinical relevant subtypes including Triple-negative, Luminal A, Luminal B, HER2 positive, COX-2 negative and COX-2 positive.

Design We followed 84,602 postmenopausal women, free of cancer in 1980, until June 2008. During this time, we prospectively collected data on analgesic use, reproductive history, and other lifestyle factors every two years. A Cox proportional hazard model was used to estimate multivariable relative risks (RRs) and 95% confidence intervals (CIs) by adjusting for age, reproductive factors and other important risk factors for breast cancer.

Results A total of 4,754 pathologically confirmed incident invasive breast cancer cases were identified during 18 years of follow-up. Comparing with never use of aspirin, multivariable relative risks of regular aspirin use (at least 2 tablets/wk) for more than 20 years were 0.91 for overall breast cancer (95%CI: 0.81-1.01, P trend = 0.16), 0.90 for ER+PR+ breast cancer (95%CI: 0.77-1.06, P trend = 0.17), 1.03 for ER+PR- breast cancer (95%CI: 0.76-1.38, P trend = 0.94), and 0.92 for ER-PR- breast cancer (95%CI: 0.68-1.23, P trend = 0.98). Results did not vary substantially by past or current use, or frequency (days per week) of use. Use of other NSAIDs and acetaminophen also was not significantly associated with risk of breast cancer. Results were not changed after mutual adjustment for each of the drug. In addition, results were not modified by body mass index, alcohol consumption, family history of breast cancer, or postmenopausal hormone use. Further, no substantial associations were noted for other subtypes of breast cancer (ER-PR+, Triple-negative, Luminal A, Luminal B, HER2 positive, COX-2 negative and COX-2 positive).

Conclusions Although a small reduction in breast cancer risk cannot be excluded, our study suggest that use of aspirin, other NSAIDs, and acetaminophen was not importantly associated with incidence of postmenopausal breast cancer overall or other specific breast cancer subtypes.

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 3724. doi:10.1158/1538-7445.AM2011-3724
Abstract 3729: Global and longitudinal analyses of demographic and clinico-pathological characteristics of triple-negative breast tumor tissues
Authors: Brown Tony , Moskalik Aaron , Pande Nupur , Peters Elizabeth H. , Sullivan Stephanie , Cohen Dalia
Published: June 2012

Patients exhibiting triple-negative (ER-negative, PR-negative, HER2-) breast cancer pose a challenge to healthcare providers due to its relatively poor prognosis, aggressive tumor reoccurrence, and lack of targeted therapies. One critical resource that accelerates the molecular and pathophysiology of the triple-negative breast cancer subtype is the analyses of retrospectively collected, verified biospecimens. Unfortunately, limited availability of adequately annotated biospecimens as well as inconsistencies in clinical data that contradict pathology diagnosis can cause ambiguity in understanding demographic and clinico-pathological characteristics in patients. In this study, we systematically examined high quality-controlled and independently molecular and pathology verified breast tumor tissues collected from patients cases in Eastern Europe, Asia, and North America diagnosed with breast cancer between 2001 and 2010 to determine age-based distributions among pre-menopausal, menopausal, and post-menopausal women with triple-negative breast cancer. We defined the immunohistochemical (IHC) surrogates for the triple-negative subtype and assessed a total of 7,636 patients including 1,700 pre-menopausal patients (22%), 4,233 post-menopausal patients (55%), and 771 menopausal patients (10%). The average RNA RIN score was 8.3 across all samples. The median age at diagnosis was 56 (range 20-91 years). Results indicated that there was a decrease in ER expression with donor age, little to no change in PR or HER2 expression, and a progressively decreasing frequency of triple-negative with donor age. Our study demonstrates that independently verified and quality controlled biospecimens enable global and longitudinal analysis of demographic and clinico-pathological characteristics of patients.

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 3729. doi:10.1158/1538-7445.AM2011-3729
Abstract 3760: Associations of risk variants for 20 different malignancies with colorectal cancer in a multiethnic population
Authors: Cheng Iona C. , Caberto Christian , Wilkens Lynne R. , Schumacher Fredrick R. , Kolonel Laurence N. , Henderson Brian E. , Stram Daniel O. , Haiman Christopher A. , Marchand Loic Le ,   PAGE Consortium
Published: June 2012

Familial clustering has been reported across different cancer sites, suggesting that cancer may be considered a broad phenotype with genetic risk factors that are shared across different malignancies. We, as part of the Population Architecture using Genomics and Epidemiology (PAGE) Study, explored the hypothesis that risk variants established for cancers at any site influence the risk of colorectal cancer. We selected 160 SNPs associated with 20 different cancer sites, including colorectal cancer, from genome-wide association studies published through May, 2009. These SNPs were genotyped in a nested case-control study of 2,077 colorectal cancer cases and 2,685 controls from five racial-ethnic groups (African American, European American, Latino, Japanese American, and Native Hawaiian) within the Multiethnic Cohort. Odds ratios and 95% Confidence Intervals were estimated by unconditional logistic regression to evaluate the association between SNPs and colorectal cancer risk, adjusting for age, sex, and race/ethnicity. We found that 22 risk variants were associated with colorectal cancer risk with P<0.05: one acute lymphocytic leukemia SNP (ALL); one basal cell carcinoma SNP; two breast cancer SNPs; one chronic lymphocytic leukemia (CLL) SNP; four colorectal cancer SNPs (rs6983267, rs4939827, rs16892766, and rs10505477); five lung cancer SNPs; one nasopharyngeal cancer SNP; one neuroblastoma SNP; two pancreatic cancer SNPs; and four prostate cancer SNPs. With more associations than would be expected by chance (8 =160 tests × 0.05), these results suggest robust significant associations. The strongest association (P<0.0001) was for a colorectal cancer risk variant at chromosome 8q24 (rs10505477). Other notable associations (P<0.01) and their originally associated cancer site, excluding known colorectal cancer risk variants (rs6983267, rs4939827, and rs4242382), include rs17483466 (ACOXL) for CLL (P=0.003), rs4242382 (8q24) for prostate cancer (P=0.004), rs10994982 for ALL (P=0.0078), and rs4975616 (TERT-CLPTM1L) for lung cancer (P=0.0079). Future analysis will include approximately 3,600 additional colorectal cancer cases and controls from the Women's Health Initiative (WHI), Arthrosclerosis Risk In Communities Study (ARIC), and the Cardiovascular Health Study (CHS) as part of the PAGE initiative. In conclusion, these results may provide important insights into the biological pathways that are shared across different cancer sites.

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 3760. doi:10.1158/1538-7445.AM2011-3760
Abstract 3764: Genome-wide association study identifies potential susceptibility loci associated with differential response to environmental insults in sporadic Chinese CRC patients
Authors: Cheah Peh Yean , Thean Lai Fun , Koh Woon Puay , Teo Yik Ying , Yuan Jian-Min , Teoh Mei Lin , Li Hui Hua , Wong Yu Hui , Koh Poh Koon
Published: June 2012

To date, genome-wide association studies (GWAS) have identified fourteen SNPs that influence risk of developing colorectal cancer (CRC) in Caucasians. None of these tagging SNPs is within coding region of a candidate gene. Only five of these SNPs were associated with CRC risk in Chinese population. We aimed to perform GWAS to identify new susceptibility loci associated with differential response to environmental insults in sporadic Chinese CRC patients. A total of 1,000 cases (aged 50 years or more) and 1,000 age- and gender-matched healthy controls were genotyped with Affymetrix SNP 6 array, a newer microarray platform which includes the ability to interrogate structural variants. Over 90% of the arrays have call rate of 99% or more. Thirty five arrays with call rate less than 97% and either null QC or negative QC (enzyme) values were replicated. Principal component analysis performed together with 270 HapMap and 92 Singapore Genome Variant Project (SGVP) samples indicate that there is no population substructure. Sixteen outliers were removed. SNPs with call rate less than 99% and minor allelic frequency less than 0.1, and control SNPs not in Hardy-Weinburg equilibrium were filtered. Subsequent quantile-quantile plot shows no evidence of allelic test statistic inflation. Twelve new chromosomal regions with SNPs having -log10 p-values greater than four and good clustering patterns were identified via Manhattan plot. Linkage disequilibrium (LD) analysis was performed to ascertain the LD blocks around these SNPs. SNPs in three chromosomal regions within the vicinity of promising CRC candidate genes were prioritized and currently being validated in a replication panel consisting of another 2,000 cases and controls.

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 3764. doi:10.1158/1538-7445.AM2011-3764
Abstract 3768: Colorectal cancer risk is not associated with increased levels of homozygosity in a population from Saudi Arabia
Authors: Siraj Abdul K. , Khalak Hanif , Al-Rasheed Maha , Sultana Mehar , Bavi Prashant , Al-Sanea Nasser , Abduljabbar Alaa , Homoud Samar Al , Ashari Luai , Al-Dayel Fouad , Uddin Shahab , Alkuraya Fowzan S. , Al-Kuraya Khawla S.
Published: June 2012

Autozygosity is a term used to denote the presence of two identical haplotypes that are derived from an ancestor shared by both parents, so it essentially represents a special type of homozygosity. Runs of homozygosity (ROH) in the genome is a measure of the extent of autozygosity and is directly correlated with the extent of inbreeding. While the role of ROH in unmasking recessively acting mutations is well established in Mendelian genetics, much less is known about their contribution to more complex disorders such as cancer. Recently, it has been suggested that ROH may contribute to the risk of colorectal cancer (CRC) perhaps through the unmasking of a recessively acting CRC-predisposing mutations in one or more genes. However, this observation could not be replicated. In this study, we examine the role of homozygosity in the CRC risk by asking four specific questions. First, do CRC patients have enrichment for ROH in particular chromosomal regions compared to controls? Second, do CRC patients have longer ROH compared to controls? Third, is there a particular SNP that is more likely to be homozygous in CRC patients compared to controls? And fourth, are CRC patients more inbred than controls? By comparing 48 Saudi CRC patients to 100 ethnically matched controls, all processed on Affy 250SytI SNP Chip platform and analyzed by Partek, we found that the answer is no to all these four questions. We note here that this is the first study to address these questions in an inbred population so the negative results in our study carry more significance than what has been previously reported in the literature. We also note that our subgroup analysis of patients with MSI-positive tumors compared to other groups did not significantly change our results. While these results do not rule out the potential presence of recessively acting CRC-predisposing genes in a small percentage of patients that our relatively small sample size could not capture, they do suggest that such genes are unlikely to account for the disturbingly high incidence of CRC in our consanguineous population and that future research should consider other mechanisms.

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 3768. doi:10.1158/1538-7445.AM2011-3768
Abstract 3791: MAPK p38 is the link between autophagy and lipid metabolism in colorectal cancer cells
Authors: Salomé Paillas
Published: June 2012

Autophagy is an intracellular degradation system that delivers cytoplasmic constituents to the lysosome. Recent progress has demonstrated that autophagy plays a wide variety of physiological and pathophysiological roles, notably in cancer.

We have showed previously, that the p38 pathway was involved in Irinotecan sensitivity of colon cancer cells p53WT.

p38 being involved in the induction of autophagy we wanted to determine its role in the processes of autophagy in presence and in absence of p53 in colorectal cancer cells.

Our results show that the overexpression of the constitutively active p38alpha isoform in HCT116 p53KO cells leads to a massive autophagy, accompanied with a block in G2 / M. The induction of autophagy in this context doesn't leads to the cellular death but rather to survival. The autophagy induction by p38 overexpression in HCT116 was weaker, indicating that the lack of p53 increase p38 role in autophagy induction.

Furthermore, we have shown that autophagy induced by p38 comes along with an increase of the lipid metabolism by activating a key enzyme of the synthesis of fatty acids.

We demonstrate here, that the role of p38 in the induction of autophagy depends on the presence of p53 and that p38 would be the link between the increase of the lipid metabolism and the induction of autophagy that lead to survival.

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 3791. doi:10.1158/1538-7445.AM2011-3791
Abstract 3800: HES6 gene is a strong glioma biomarker and a key transcriptional regulator needed for cancer cell growth
Authors: Haapa-Paananen Saija M. , Kiviluoto Santeri , Waltari Marika , Puputti Marjut , Mpindi John-Patrick , Kohonen Pekka , Tynninen Olli , Haapasalo Hannu , Joensuu Heikki , Perälä Merja , Kallioniemi Olli
Published: June 2012

Malignant glioma is the most common brain tumor with 16,000 individuals diagnosed annually in the United States. We performed a systematic large-scale transcriptomics data mining study of 9,783 Affymetrix samples from the GeneSapiens database (www.genesapiens.org) to systematically identify genes that are most glioma-specific. We searched for genes that were highly expressed in 322 glioblastoma multiforme tissue samples and 66 anaplastic astrocytomas as compared to 425 samples from histologically normal central nervous system. Transcription cofactor HES6 (Hairy and Enhancer of Split 6) emerged as one of the most glioma-specific genes. Immunostaining of a tissue microarray showed HES6 expression in 335 (98.8%) out of the 339 glioma samples. HES6 was also expressed in endothelial cells in normal and glioma tissues. Recurrent grade 2 astrocytomas and grade 2-3 oligodendrogliomas showed higher levels of HES6 immunoreactivity than the corresponding primary tumors. Functional studies implied a critical role for HES6 in supporting survival of glioma cells, as evidenced by reduction of cancer cell proliferation and induction of Caspase 3/7 activity after HES6 silencing in A172 and LN405 cells. The biological role and consequences of HES6 silencing and overexpression was explored with genome-wide analyses, which implicated a role for HES6 in p53, c-myc, and NF-κB transcriptional networks. We conclude that HES6 has a critical role in sustaining glioma cell growth, survival, migration and possibly angiogenesis. HES6 is a potential therapeutic target and biomarker for glioma.

This work was supported by funding under the 6th FP of the European Union, Project RIGHT (LSHB-CT-2004-005276), and the Academy of Finland.

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 3800. doi:10.1158/1538-7445.AM2011-3800
Abstract 3805: Novel chemosensitive single-nucleotide polymorphism markers to targeted regimens in metastatic colorectal cancer
Authors: Kim Jin Cheon , Kim Seon Young , Cho Dong Hyung , Ha Ye Jin , Choi Eun Young , Kim Chan Wook , Roh Seon Ae , Kim Tae Won , Ju Hyoung Seok , Kim Yong Sung
Published: June 2012

Purpose: Methods for predicting individual responsiveness to targeted chemotherapy are urgently needed, considering the frequent resistance and extremely high cost of such therapies.

Experimental Design: A chemosensitive single-nucleotide polymorphism (SNP) discovery schema is presented that utilizes genome-wide SNP screening with a human SNP array, an in vitro chemosensitivity assay in 118 colorectal cancers, clinical association analysis in 98 patients who had received metastatic chemotherapy, and biological utility assessment using cell viability assays of transfected colorectal cancer (CRC) cells.

Results: Eleven SNPs related to bevacizumab and cetuximab sensitivity were initially chosen during screening. Response rates and survival periods revealed that patients carrying specific alleles of the SNPs ANXA11 rs1049550, LINS1 rs11247226, or ITGA3 rs2230392 were chemosensitive to bevacizumab regimens, and patients with DFNB31 rs2274159, LIFR rs3729740, or ISX rs361863 were chemosensitive to cetuximab regimens. Cytotoxicity analyses using MTT and caspase-3 assays showed that all RKO and HCT116 CRC clones transfected with the reference alleles of LIFR rs3729740 and ISX rs361863 (G and C, respectively) were more sensitive to cetuximab regimens than those transfected with the substitution alleles.

Conclusions: Chemosensitive SNP markers were identified using a novel three-step process. The chemosensitive markers LIFR rs3729740 and ISX rs361863 will hopefully predict responsive patients to cetuximab regimens, although further validation is needed in large cohorts.

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 3805. doi:10.1158/1538-7445.AM2011-3805
Abstract 381: Light-absorbing nanoparticles induce cell death in esophageal cancer cells
Authors: Li Yan , Gobin Andre M. , Dryden Gerald W , Xiao Deyi , Zhang Guandong , Martin Robert C.
Published: June 2012

Background Light-absorbing nanoparticles that are nontoxic to biological tissue possess the potential for targeted killing of cancer cells while causing minimal collateral damage to adjacent healthy tissue. Gold nanoparticles with near infrared (nIR)-absorbing light properties are ideal for this application. Aim: This study is to investigate the photo-thermal ablation effect of nIR-absorbing gold nanoparticles coated with a natural polymer (chitosan) on esophageal cancer cells as well as benign cells. Methods: Three esophageal cell lines used are human squamous esophageal epithelium cell line (Het-1A), Barrett's epithelium cell line (BAR-T), and the esophageal adenocarcinoma cell line (OE-19). Furthermore, the uptake of gold nanoparticles in the benign esophageal tissue and the response to laser exposure was also valuated in a rat esophagoduodenal anastomosis (EDA) model. Three cell lines were treated with gold nanoparticles (OD=25) then exposed to laser at a wavelength of 817 nm and power density of 3W/cm2 for 3 min. The esophageal mucosa of rats was sprayed with gold nanoparticles 24 hours befor laser application. After treatment, the rats were exposed to laser at a wavelength of 817 nm and power density of 4.15W/cm2 for 3 min. The loading of gold nanoparticles was detected by silver staining. Results: The most OE-19 cells loaded with gold nanoparticles and were completely ablated after nIR light exposure. However, most Het-1A cells and BAR-T cells didn't load on any gold nanoparticles, and they remained viable after nIR light exposure. Normal esophageal epithelium and Barrett's mateplastic epithelium show low affinity to gold nanoparticles, however, the focal region with more loaded gold nanoparticles was ablated after nIR light exposure. Conclusion: 1) Chitosan coated gold nanoparticles undergo targeted uptake by OE-19 cells, with a much higher affinity compared to benign cells. 2) Photo-thermo-ablation by nIR light exposure causes a directed cancer cell-killing effect based on preferential gold nanoparticle uptake. 3) Further study is ongoing to investigate methods of enhancing peak uptake of gold nanoparticles in the EDA rat model, to provide an optimal model for laser ablation of esophageal cancer.

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 381. doi:10.1158/1538-7445.AM2011-381
Abstract 3819: Matrix metalloproteinase-1 (MMP1) is an important marker of oral squamous cell carcinoma
Authors: Stott-Miller Marni , Houck John , Lohavanichbutr Pawadee , Schwartz Stephen M. , Upton Melissa P. , Chen Chu
Published: June 2012

The matrix metalloproteinase genes have frequently been identified as important in cancer development and progression, including in oral squamous cell carcinoma (OSCC). These proteases are believed to cause degradation of the extracellular matrix and basement membranes and therefore to play a key role in tumor metastasis, cell migration, cancer cell growth, and angiogenesis. We compared MMP1 expression levels in primary OSCC (n=119) with expression levels in normal oral epithelium from control subjects (n=35), as measured by Affymetrix U133 2.0 Plus GeneChip arrays, adjusting for age and sex. MMP1 was expressed more than 200-fold higher in OSCC compared with normal control epithelium. qRT-PCR results on 30 of the cases and 22 of the controls confirmed the substantial differential expression of MMP1, with a difference in mean threshold cycle (Ct) value for cases versus controls of 9.85 (95% CI, 8.74-10.97, p<0.001). In an independent testing set of 48 OSCC tissue samples and 10 control samples, MMP1 had exceptional discriminatory power to separate cases from controls (AUC=100%). This result was further validated in an external data set from Gene Expression Omnibus (GEO; GSE6791) of 42 OSCC cases and 14 controls (AUC=98.4%; 95% CI, 95.6%-100%). The mean of the log2 MMP1 expression levels on the Affymetrix arrays for 45 normal oral epithelium was 3.49 (95% CI, 3.21-3.78), 17 dysplasia tissue samples had mean expression of 6.69 (95% CI, 5.21-8.17), 108 stage T1/T2 cases had a mean of 10.93 (95% CI,10.49-11.37), and the mean of 57 stage T3/T4 cases was 11.38 (95% CI,10.81-11.95). These results suggest that MMP1 is increasingly dysregulated during the process of oral carcinogenesis. The capacity of MMP1 to identify OSCC provides support for further investigation into this enzyme as a potential marker for OSCC development and/or progression.

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 3819. doi:10.1158/1538-7445.AM2011-3819
Abstract 385: Monocyte chemoattractant protein-1 (MCP-1)/CCL2 produced by non-tumor cells in tumor stroma promotes lung metastasis of 4T1 murine breast cancer cells
Authors: Yoshimura Teizo , Zack Howard O. M. , Matsukawa Akihiro , Chen Keqiang , Liu Ying , Wang Ji Ming , Oppenheim Joost J.
Published: June 2012

MCP-1 is a chemokine that plays an important role in the initiation and progression of cancer by attracting immunosuppressive macrophages. A wide variety of tumor cell lines produce MCP-1 in vitro; thus, tumor cells are considered to be the main source of this chemokine. In the present study, we examined whether MCP-1 deficiency in non-tumor cells present in tumor stroma affects the growth and metastasis of the 4T1 mouse breast cancer cells implanted into the mammary pad of wild-type (WT) or MCP-1-deficient mice. Tumors grew at a similar rate in both WT and MCP-1-deficient mice at the injected sites; however, the lung metastases were markedly reduced in MCP-1-deficient mice. Serum MCP-1 levels increased after 4T1 cell implantation in WT, but not in MCP-1-deficient mice. High levels of MCP-1 mRNA were detected in the mammary tumors of WT, but not of MCP-1-deficient mice, indicating that tumor cells were not the main source of MCP-1. In tumors of MCP-1-deficient mice, the infiltration of macrophages was reduced and the expression of cytokines, including IFN-γ, IL-4, IL-10 and IL-13, was elevated. Transplantation of MCP-1-deficient bone marrow cells into WT mice did not affect the number of lung metastasis, whereas transplantation of WT bone marrow cells into MCP-1-deficient mice significantly increased the lung metastasis. Finally, culture supernatants of 4T1 cells induced MCP-1 production in inflammatory macrophages. These results suggest that MCP-1 produced by non-tumor cells in tumor stroma, both bone marrow-derived and non-bone marrow-derived, facilitates lung metastasis of 4T1 cells by recruiting immunosuppressive macrophages. Thus, the inhibition of MCP-1 may be useful to reduce lung metastasis of breast cancer cells.

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 385. doi:10.1158/1538-7445.AM2011-385
Abstract 3852: Identification and functional characterization of T-cell lymphoma invasion and metastasis 2 (TIAM2) in neuronal cells
Authors: Chen Jia-Shing , Sun H. Sunny
Published: June 2012

T-Cell Lymphoma Invasion and Metastasis 2 (TIAM2) gene was identified as the homolog for human TIAM1 gene, a Rac-specific guanine nucleotide exchange factor (GEF), and classified as a member of TIAM GEF subfamily. Although functions of TIAM family proteins have been extensively studied, the characteristics of human TIAM2 remain largely unknown. In this study we systematically investigate the cellular function of TIAM2. Two mRNA isoforms of TIAM2, TIAM2S and TIAM2L, were detected and showed distinct expression patterns in various human tissues. Although both mRNAs are highly expressed in many human tissues, no TIAM2L protein can be detected in any of the examined human tissues and cell lines. In addition, TIAM2S protein only expressed in neuron of restricted regions in the human brain. Overexpression of TIAM2S-GFP fusion protein revealed a cytosolic distribution in both HeLa and NIH3T3 cells. Using LC/MS/MS and enzymatic analysis, TIAM2S was shown to be an O-linked glycoprotein. As transiently transfected TIAM2S-fusion protein neither translocate onto the cell membrane, nor exhibit GEF activity to RhoGTPases in vivo, these data suggested human TIAM2S is not a classical GEF and its functional role in normal brain may differ from other TIAM members.

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 3852. doi:10.1158/1538-7445.AM2011-3852
Abstract 3859: Defining groupings of oropharyngeal squamous cell carcinomas by expression patterns of human papillomavirus oncogenes
Authors: Laborde Rebecca R. , Olsen Steven D. , Wang Vivian , Garcia Joaquin J. , Graham Rondell , Berres Michael W. , Halling Geoffrey C. , Olsen Kerry D. , Moore Eric J. , Kasperbauer Jan L. , Tombers Nicole D. , Smith David I.
Published: June 2012

There is considerable clinical interest in the role that human papillomavirus (HPV) plays in the development of oropharyngeal cancers. A variety of assays are currently available to stratify patients based on HPV status prior to treatment. The most frequently utilized are PCR and In Situ hybridization (ISH) to detect the presence of HPV DNA sequences. The increasing clinical interest in determining HPV patient status relates to compelling evidence that a subset of patients with HPV positive tumors have better clinical outcomes. Evidence also suggests that patients with HPV positive tumors should be further divided into groupings based on levels of viral activity and that this may represent additional clinical information necessary to develop appropriate treatment plans. The most frequently applied assay to identify HPV activity in a patient sample is to detect the over-expression of the surrogate protein marker p16 by Immunohistochemisty (IHC), which has been shown to increase expression in response to HPV active infections in oropharyngeal cancer. An alternative technique to quantitatively detect the presence and level of transcriptional activity of HPV in tissue samples is to measure the expression of oncogenes E6 and E7. We have analyzed the fresh frozen tissue samples corresponding to matched tumor, normal and metastatic patient tissues using a quantitative PCR (qPCR) approach to determine the expression of HPV oncogenes and p16 gene expression. We observed that there are three groups of oropharyngeal tumors when grouped by HPV16 E6 and E7 gene expression. The first are the most rare and are tumors that contain no HPV sequences or expression of the E6 and E7 ocogenes. The second are tumors that contain HPV DNA sequences but very low expression of E6 or E7 and the third are tumors that contain HPV DNA sequence and robust expression of E6 and E7. Additionally, we found variable correlation between HPV16 E6 and E7 expression and p16 gene expression, suggesting that factors other than HPV oncogene expression may influence the expression of p16. Our results would indicate that the best assays for quantifying the activity of an HPV infection in oropharyngeal cancers would be those optimized to accurately measure either E6 and E7 transcripts or their protein products directly out of paraffin-embedded tissues.

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 3859. doi:10.1158/1538-7445.AM2011-3859
Abstract 3867: EGFR genotype impacts expression of chemotherapeutic target biomarkers in NSCLC
Authors: Liu H Eugene , Liu Jacqueline M. , Yu Ming-Chih , Bai Kuan-Jen , Lan Chieh
Published: June 2012

BACKGROUND: Non small cell lung cancer EGFR mutations are prevalent in the East Asian population, especially in female non-smokers, the L858R and del 747-753 in exons 18-21 of the EGFR tyrosine kinase domains are activating mutations with increased sensitivity to the EGFR tyrosine kinase inhibitors (TKIs) gefitinib and erlotinib. EGFR gene amplification also increases susceptibility to EGFR TKIs as has been reported in the IDEAL / INTACT non small cell lung cancer (NSCLC) trials. Our study evaluated chemotherapeutic target biomarkers in Chinese NSCLC patients with varying EGFR genotypes. METHODS: Seventy NSCLC patients with known EGFR genotype received chemotherapy and EGFR TKI therapy with known response and survival, their pathology specimen were retrospectively retrieved for evaluation. EGFR and KRAS were sequenced for mutations and deletions. Fluorescent in situ hybridization (FISH) was performed for EGFR genes. Immunohistochemical analyses were performed for topoisomerase 1, c-myc, RRM-1, class III beta tubulin, topoisomerase IIα, p53, thymidylate synthase, and ERCC-1. The percentage of nuclear-stained malignant cells were recorded for T IIα, T1, p53, ERCC-1, c-myc, whereas for TS, tau, RRM-1, class III beta tubulin were cytoplasmic stains, only positive or negative was reported, eventually only 63 patients had complete data for evaluation. RESULTS: Significant increase in overall survival was found in females (p=0.002), non-smokers (p=0.005), younger age (p=0.001), less advanced staging (p=0.048), EGFR mutation (p=0.038); but not with KRAS genotype, or expression of any chemotherapeutic markers. Patients in this study were consecutive patients, 60-70% received treatment. When EGFR mutant tumors were compared to EGFR wildtype tumors, the mutant tumors had lower KRAS mutation (4% vs 19%, p=0.1238), more were ERCC1- (76% vs 63%, p=0.2925), more topoisomerase 1 + (28% vs 17%, p=0.3719), less RRM1+ (24% vs 35%, p=0.4142), more TS- (93% vs 86%, p=0.4478), lower class III beta tubulin (45% vs 57%, p=0.4515). CONCLUSIONS: A difference in expression of chemotherapeutic target markers has been detected in differing EGFR genotypes, although its significance has been dampened by our small patient population, and larger patients groups will be needed to evaluate the impact of different chemotherapies on survival of the different EGFR genotype tumors.

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 3867. doi:10.1158/1538-7445.AM2011-3867
Abstract 387: Prostate cancer cell invasion is stimulated by cross communication with human monocyte-lineage cells
Authors: Lindholm Paul F. , Sivapurapu Neela , Jovanovic Borko , Crawford Susan E.
Published: June 2012

Tumor-associated macrophages (TAMs) produce factors that may stimulate tumor growth, angiogenesis and progression. In this study, co-cultures with monocyte-derived THP-1, U-937 cells or normal peripheral blood monocytes stimulated PC-3, DU145 and LNCaP prostate cancer cell basement membrane invasion. This study investigated the interactions between monocytes and prostate cancer cells leading to increased cancer cell invasion. The PC-3 prostate cancer cells expressed high levels of IL-6 and IL-8, at similar levels to co-cultures with human U-937 monocyte-derived cells. However, the co-culture supernatants showed increased MCP-1 levels, up to 5 ng/mL, compared with prostate cancer cells cultured alone with MCP-1 less than 10 pg/mL. During the co-cultures, MCP-1 was first detected at 8 hours and increased over 3 days. Co-culture of U-937 or THP-1 monocytes with either DU145 or LNCaP cells also increased MCP-1 expression significantly above cancer cell or monocyte cultures alone. When cultured separately, the monocyte-lineage cells produced more MCP-1 than the cancer cells; however, MCP-1 expression increased more than 10-fold in the co-cultured prostate cancer cells. Addition of purified MCP-1 and IL-8 stimulated prostate cancer cell invasion to levels similar to the co-cultures. Further, addition of anti-MCP-1 neutralizing antibodies inhibited prostate cancer cell invasion when compared with control antibodies. Co-culture with U-937 cells also stimulated NF-κB DNA binding activity in the prostate cancer cells. Transfection of the prostate cancer cells with dominant negative IκBα or treatment of the co-cultures with Bay11-7082 or Lacticystin inhibited NF-κB DNA binding and prostate cancer cell invasion. These experiments demonstrate that human monocyte-derived cells can cross-communicate with prostate cancer cells leading to increased cancer cell invasion. These studies have potential to identify treatments to target MCP-1 and NF-κB activity to inhibit monocyte-induced cancer cell invasion and progression. Support by Northwestern University Prostate S.P.O.R.E. P50CA90386 and CDMRP DAMD-17-02-1-0162

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 387. doi:10.1158/1538-7445.AM2011-387