bluuebunny/crossref_metadata_2025_split huggingface dataset:
EGFR expression in invasive anal carcinoma.
Authors: Paliga A. A. , Onerheim R. , Gologan A. , Spatz A. , Vuong T. 
Published: February 2017

412 Background: Squamous cell anal carcinoma (SCAC) treatment remains unchanged since the institution of chemoradiation over 4 decades ago. Epidermal growth factor receptor (EGFR) is a protein often expressed in aggressive cancers and that is the target of the monoclonal antibody: cetuximab. Concurrent cetuximab and radiation has been particularly effective treating squamous cell carcinoma of the head and neck. Like head and neck cancer, anal cancer is an epithelial tumor of the alimentary tract that is radioresponsive and is associated with HPV infection. The rarity of this cancer limits its evaluation for biological markers. This study set out to thoroughly characterize EGFR expression by immunohistochemistry in 101 invasive SCAC tissue samples. Methods: One hundred and one pretreatment paraffin embedded invasive SCAC biopsies, obtained from the Montreal area between 1999 and 2009, were tested for EGFR expression by immunohistochemistry. All samples were confirmed to harbor invasive anal carcinoma on H&E slide preparations. Corresponding cancerous areas were identified on paraffin tissue blocks and cut out for tissue microarray analysis. Samples were immunostained with an EGFR antibody (clone SPM 341) on the Discovery XT Autostainer (Ventana), and staining was assessed by light microscopy by two pathologists. A semiquantitative combination score combining staining intensity with the percent of cells staining gave a final score: just detectable or weak (1+); moderate (2+); strong/intense (3+). Results: Of 101 patient biopsies, 82 samples had sufficient material for interpretation. Of these samples, 72/82 (90%) stained positive for EGFR, while 41/82 (50%) samples displayed at least moderate to strong staining. Conclusions: This is the largest cohort of SCAC tissue samples tested to date for EGFR expression and it confirms that the vast majority of invasive SCAC overexpress EGFR. EGFR likely plays a role in anal cancer tumor-genesis and progression. Testing of EGFR inhibitors in this patient population is justified. No significant financial relationships to disclose.
Use of gene expression profiling to determine prognosis and therapeutic targets for patients with appendiceal carcinoma.
Authors: Kim M. K. , Blazer D. G. , Stewart J. H. , Guy C. , Shen P. , Levine E. , Hsu S. D. 
Published: February 2017

418 Background: Appendiceal carcinomas are rare neoplasms that often present as peritoneal surface malignancies. The tumors are treated similarly to primary colorectal metastases, despite having different biological environments. The aim of this study is to utilize genomic analyses to understand the underlying biology of appendiceal carcinoma in order to better guide prognosis and therapy. Methods: Forty-one metastatic human tumors to the peritoneum (26 appendiceal carcinoma, 15 colorectal carcinoma) were subjected to global gene expression analysis. Unsupervised hierarchical clustering was used to differentiate phenotypes based on differential gene expression. Phenotypes were further analyzed to find differences in oncogenic pathway deregulation and findings were validated using gene specific inhibitors in vitro. Results: Unsupervised hierarchical clustering revealed three distinct phenotypes, two consisting of predominantly low grade appendiceal samples (Cluster 1 and Cluster 2) and one consisting of predominantly colorectal samples (Cluster 3). Cluster 1 consisted of patients with good prognosis and Cluster 2 consisted of patients with poor prognosis (p = 0.046). Further characterization of Cluster 1 and Cluster 2 with oncogenic pathway signatures identified increased deregulation of the EGFR pathway in the poor prognosis cluster. Similarly, colon cancer cell lines with the poor prognosis phenotype were found to be more sensitive to gefitinib (EGFR inhibitor) (p = 0.038). Conclusions: The use of geneexpression profiling can be used to identify prognosis and novel therapeutic targets for patients with appendiceal carcinoma. These methods can serve as models to understand the biology of appendiceal carcinoma and to assist in the design of future trials for patients with appendiceal cancer. No significant financial relationships to disclose.
The significance of HPV status in patients with anal cancer: A comparative technical analysis.
Authors: Doll C. M. , Roldán G. B. , Klimowicz A. , Gustafson K. , Petrillo S. K. , Chan A. K. , Lees-Miller S. P. , Magliocco A. M. 
Published: February 2017

425 Background: HPV status is an important prognostic factor for outcome in head and neck cancers, with improved survival noted in HPV-positive vs -negative tumors. However, this effect has not been studied in anal cancer. Challenges exist in defining reliable and methods of detecting the presence of high-risk HPV virus or the expression of HPV-related proteins. Purpose: To determine p16 expression and HPV16 status, in association with HPV subtyping, in pre-treatment anal cancers, and to correlate marker status with clinical outcome. Methods: This retrospective study evaluated anal cancer patients treated between 1992-2005 with definitive RT or CRT at a single institution (Tom Baker Cancer Centre, Calgary, Canada). HPV subtyping, p16 protein expression via conventional immunohistochemistry (IHC) and automated quantitative IHC (AQUA), and HPV16 in-situ hybridization (CISH) was performed in a subset of patient tumors with sufficient pretreatment tumor specimen. The correlation between results using these different techniques and association of HPV markers with clinical outcome was evaluated. Results: 89 patients were identified; M:F 1:2; median age 57 years. Median tumor size was 3.5 cm. All patients were treated with radical external beam RT with curative intent; 81% received concurrent chemotherapy. Clinical CR was observed in 73% of patients at 3 months post-treatment. Median OS was 82 months. Of tumor specimens analyzed for HPV subtype, 78% (28/36) were HPV16. Using conventional IHC (DAB), 80% (28/35) of tumors over-expressed p16 (score 3). HPV16 CISH was positive in 81% (34/42). P16 AQUA™ score correlated with HPV16 subtyping, CISH for HPV16, and p16 DAB. With HPV16 subtyping as the gold standard for HPV16 infection, false positive and false negative rates were 8% and 4% with DAB, and 10% and 13% with CISH. In univariate analysis only p16 AQUA score > 244 (upper 15%) was associated with PFS (p= 0.006-95% CI 8.4 [1.8-38.6]) and OS (p= 0.013-95% CI 4.5 [1.4-15]). Conclusions: p16 expression via quantitative IHC correlates with other methods, including HPV16 subtyping, and appears more reliable for defining HPV16 status than CISH. Only quantitative IHC identified a subset of patients with worse outcome. No significant financial relationships to disclose.
Polymorphisms of thymidylate synthase as prognostic factor in rectal cancer.
Authors: Arrazubi V. , Suarez J. , Guerrero D. , Cambra K. , Gomez Dorronsoro M. L. , Arias F. , Balen E. , Teijeira L. , Vera R. 
Published: February 2017

433 Background: Neoadjuvant fluoropyrimidine-based chemotherapy (ChT) plus radiotherapy (Rt) is a standard approach for locally advanced rectal cancer. Polymorphisms of thymidylate synthase (TS), the target for fluoropyrimidines, are recognized prognostic factors in colon cancer. The aim of this study was to evaluate the prognostic value of the polymorphisms of TS in rectal cancer after neoadjuvant ChT plus Rt. Methods: We studied one-hundred consecutive patients with stage II/III rectal cancer between November 2001 and March 2009. Patients underwent surgery 6-8 weeks after neoadjuvant Rt (5,040 cGy) plus fluoropyrimidine-based ChT. DNA was extracted from paraffin embedded biopsies. TS1494del6 and 5′-28bp repeat +G/C SNP polymorphisms were determined. Results: Sixty-seven percent were men and median age was 67 years. ypT stage was: T0 9%, T1 2%, T2 27%, T3 60% and T4 2%; 32% had locoregional adenopathies. The median follow-up was 45 months and relapse occurred in 20% of patients. Polimorphisms could be determined in 98% of pt: -6bp/-6bp 10%, - 6bp/+6bp 39%, +6bp/+6bp 51% and 2R/2R 72%, 2R/3R 21%, 3R/3R 6%. The grade of pathological tumour regression was not associated with polymorphisms. Relapses occurred in 40% of patients -6bp/-6bp, 22% of patients -6bp/+6bp and 21% of patients +6bp/+6bp. The difference in disease- free survival (DFS) between the first and the third groups was stadistically significative (p=0.049). No relation between 5′-28bp repeat +G/C SNP polymorphism and DFS was found. Conclusions: Our data suggest that the TS1494del6 polimorphism may be an important prognosis factor in rectal cancer receiving neoadjuvant chemoradiotherapy. No significant financial relationships to disclose.
Contribution of the epithelial-mesenchymal (EMT) phenotype to the sensitivity of colorectal cancer cell lines to the p21-activated kinase inhibitor, PF-3758309.
Authors: Im E. , Pitts T. M. , Kulikowski G. , Tan A. , Tentler J. J. , Murray B. , Carley W. , Smeal T. , Guo C. , Eckhardt S. G. 
Published: February 2017

438 Background: Increased expression of p21–activated kinase (PAK) family proteins has been observed in a range of malignancies including those of breast, ovarian, colorectal (CRC), and pancreatic origin. PAK (PAK 1-6) proteins are critical mediators of cell proliferation, motility, transcription, and translation. In initial studies, we demonstrated that the PAK4 inhibitor, PF-3758309, exhibited robust single agent efficacy against selected CRC cell lines, with IC50s of less than 0.015 uM, whereas some cell lines were relatively resistant to the agent, with IC50s of greater than 1 uM. In subsequent gene array analyses, CRC cell lines sensitive or resistant to PF-3758309 exhibited overexpression of core genes associated with a mesenchymal or epithelial phenotype, respectively. Thus, the goal of this study was to assess the functional consequences of altering the expression of EMT-associated genes and to seek rational combination partners in CRC. Methods: CRC cell lines that were sensitive to PF-3758309 (IC50=0.015 uM) were transfected with selected short hairpin- (sh)RNAs or micro-(mi)RNAs that are known to regulate EMT. Semi-quantitative RT-PCR and immunoblotting were performed to confirm target knockdown. The transfected cell lines were then exposed to increasing concentrations of PF-3758309 to determine the functional role of these genes in conferring responsiveness to PF-3758309. Results: Sensitive CRC cell lines were transfected with shRNAs to Zeb1, vimentin, and caldesmon, genes that are associated with a mesenchymal phenotype. Interestingly, when the cell lines were exposed to increasing concentrations of PF- 3758309 they demonstrated a ′right shift′ towards a more resistant phenotype. Likewise, transfection with miRNA 200c, a known suppressor of Zeb1, resulted in a similar shift towards resistance. Conclusions: These data suggest that the EMT phenotype may play a functional role in determining CRC sensitivity to the PAK4 inhibitor PF-3758309, and in addition, a rational combination targeting the epithelial phenotype with epidermal growth factor receptor inhibitors, may be warranted. [Table: see text]
The expression of frizzled-3 receptor in colorectal cancer and colorectal adenoma.
Authors: He W. , Wong S. C. , Ma B. , NG S. S. , Lam M. Y. , Chan C. M. , Au T. C. , Chan J. K. , Chan A. T. 
Published: February 2017

444 Background: Wnt signaling is well known for its role in colorectal cancer (CRC) formation through transcriptional activities of nuclear β-catenin. Although activation of Wnt signaling depends on specific Wnt/Frizzled receptors (FZD) combinations, the specificity of the interaction and the role of FZD in that particular interaction are still unknown. Among the 10 Wnt receptors of the FZD protein family, FZD-3 is involved in neurodevelopmental abnormalities and gastric cancer carcinogenesis. However, the expression of FZD-3 in CRC is not clear. Therefore in this study, we examined the expression of FZD-3 in CRC cell lines and CRC patient tissues with various pathological stages. The information obtained will be important for us to understand the role of FZD-3 in the development of CRC. Methods: FZD-3 mRNA expression was studied in CRC metastatic SW620, primary SW480 and normal CCD18co cell lines using quantitative real-time polymerase chain reaction with primers and a Taqman minor grove binder probe (Applied Biosystems, Foster City, USA). Moreover, paraffin-embedded specimens of 40 CRC patient tissues, 25 colorectal adenoma (CA) tissues were retrieved from the Department of Pathology, Queen Elizabeth Hospital, Hong Kong Special Administrative Region for FZD-3 immunostaining using an anti-FZD-3 antibody (Catalog no: MAB1001, R&D systems Inc., Minneapolis, USA) in an automatic Ventana Benchmark XT immunostainer (Ventana Medical Systems Inc., Tucson, USA). Results: FZD-3 mRNA was up-regulated in metastatic SW620 cell line (fold-change: 622) and in primary SW480 cell line (fold-change: 820) when compared to that in normal CCD18co cell line. Furthermore, immunostaining showed that FZD-3 protein was expressed in 100% (40/40) of CRC specimens and 84% (21/25) of CA specimens. Detailed analysis showed that FZD-3 protein was significantly up-regulated in CRC, CA when compared to their adjacent normal colorectal epithelial tissues (p < 0.0005, Wilcoxon matched pairs test). Conclusions: This study provided evidence that FZD-3 is involved in CRC carcinogenesis and it is a potential therapeutic target in CRC. No significant financial relationships to disclose.
Clinical outcome in patients with metastatic colorectal cancer harboring KRAS p.G13D mutation treated with cetuximab.
Authors: Bando H. , Yoshino T. , Shinozaki E. , Yuki S. , Nishina T. , Kadowaki S. , Yamazaki K. , Tsuchihara K. , Fujii S. , Yamanaka T. 
Published: February 2017

448 Background: Metastatic colorectal cancer patients with KRAS codon 12 or 13 mutated tumors are presently excluded from treatment with cetuximab (Cmab). On the other hand, a few patients who have mutated KRAS status occasionally respond to Cmab. The tumors of those patients predominantly had codon 13 mutation, and all codon 13 responder have mutation of p.G13D. We now compared the efficacy of Cmab among patients with p.G13D- mutant, other KRAS mutant and KRAS wild-type colorectal cancer. Methods: The patients from 9 Japanese institutions were retrospectively collected and analyzed. All patients were refractory to fluoropyrimidine, oxaliplatin and irinotecan, and were treated with Cmab and irinotecan combination regimen. Response rate (RR), progression-free survival (PFS) and overall survival (OS) were calculated respectively according to KRAS status. Results: Ninety four patients were treated with combination therapy. Among 94 cases, 7 cases were p.G13D-mutant KRAS, 23 cases were other mutant KRAS and 63 cases were wild-type KRAS. Baseline characteristics by each subset were well-balanced. While one partial response (PR) and 4 stable diseases (SD) cases were found in 7 p.G13D-mutated cases, no PR was found in other KRAS mutated cases. Median PFS of the patients with p.G13D-mutant, other KRAS mutant and KRAS wild-type were 4.5 months (95%CI 1.7-), 2.3 months (95%CI 1.9-4.3), 4.6 months (95%CI 3.5-6.5) respectively. And median OS of the patients with p.G13D- mutant, other KRAS mutant and KRAS wild-type were 9.3months (95%CI 8.5- 11.8), 7.4 months (95%CI 4.5-9.4), 12.2 months (95%CI 8.7-19.8) respectively. Although statistical significance was not found between the two mutated groups, there are trends that the patients with p.G13D-mutant may have received better clinical benefits from Cmab than the patients with other KRAS mutant. Conclusions: Cmab may have therapeutic benefit in the patients with KRAS p.G13D-mutant colorectal cancer although further evaluation is warranted. No significant financial relationships to disclose.
MicroRNA profile and KRAS status in metastatic colorectal cancer.
Authors: Valladares-Ayerbes M. , Blanco M. , Reboredo M. , Lorenzo-Patiño M. J. , Arnal F. , Haz Conde M. , Medina Villaamil V. , Santamarina Caínzos I. , Antón Aparicio L. M. , Calvo Martinez M. 
Published: February 2017

453 Background: microRNAs (miRNAs) are small non-coding RNA with regulatory functions. miRNAs expression is deregulated in cancer. Our objective is to establish a miRNA expression profile as a prognostic and predictive signature in Colorectal Cancer (CRC). In this cohort, patients with metastatic (m) CRC have been included. The expression of miRNAs have been analyzed and correlated with clinical and pathologic parameters, including KRAS status, disease free survival (DFS), and overall survival (OS). Methods: Tissue samples of mCRC were obtained as formalin-fixed paraffin-embedded (FFPE) specimens from the archives of our institution, after obtaining informed consent of patients. Genomic DNA was extracted using Qiaamp DNA mini kit (Qiagen). For the mutational testing of KRAS gene, we use the TheraScreen KRAS PCR kit (Qiagen). miRNA-enriched RNA was extracted using High Pure miRNA Isolation Kit (Roche). miRNA expression was analyzed by quantitative reverse transcription PCR (RT-qPCR) using LNA technology (Exiqon) in a LightCycler 480 (Roche) device. REST2009 (Qiagen), SPSS 19 (IBM), and Prism (GraphPad) software was used to data analysis. Results: One-hundred and forty mCRC patients have been included. To date, we have extracted DNA and performed KRAS mutational testing for 127 FFPE specimens. From these, 72 (56.7%) were wt while the remaining 55 (43.3%) were mt. We have performed a miRNA expression studies in 28 tumors. miRNAs let-7a, miR-17, miR-21, and miR-200c were analyzed. 5S rRNA and U6 snRNA were amplified as reference genes. miR-21, miR-17, and let-7a were found upregulated (p<0.05) in mCRC samples versus healthy colon. However, for all 4 miRNAs analyzed, there were no significant differences of expression between wt and mt tumor samples for KRAS gene. Conclusions: The expression of 4 miRNAs analyzed is not correlated with mutations in KRAS gene. However, 3 of 4 miRNAs analyzed were found upregulated in tumors. The results for the association of the expression of miRNAs analyzed with the response to treatment and other clinical parameters will be presented during the symposium. Supported in part by SERGAS, Xunta de Galicia, Grant PS08/77. No significant financial relationships to disclose.
In silico and ex vivo approaches identify a role for toll-like receptor 4 in the biology of sporadic colorectal cancer.
Authors: Sussman D. A. , Santaolalla R. , Clarke J. , Abreu M. T. 
Published: February 2017

460 Background: Several lines of evidence suggest that the host response to bacteria plays a role in the development of colorectal cancer (CRC). Work on innate immune signaling has shown that toll-like receptor-4 (TLR4) is required for colitis-associated neoplasia in a mouse model, suggesting that TLRs may be a critical link between colonic bacteria and the development of colon cancer. The objective of the present study was to test the hypothesis that dysregulated TLR4 expression occurs in a subset of sporadic colorectal cancers. Methods: Bioinformatic and immunohistochemical approaches were used to test the hypothesis. Publicly available, online microarray data sets of gene expression data (Oncomine and Gene Expression Omnibus) were searched for information relevant to CRC. Searches were included if data sets contained human patient data with paired samples, ≥ 20 subjects per study, and use of standardized microarray platforms. Excluded searches contained patients with genetic cancer syndromes or colitis. Associations were investigated between TLR4 expression and oncogenic pathways. Staining of human CRCs for TLR4 was performed in a consecutive series of 10 patients at the University of Miami. Intensity of staining for TLR4 was calculated using immunofluorescence and NIH's ImageJ software. 5 different areas per 10x-field were quantified per tumor. Results: Heat maps were generated from the microarray data and demonstrated an increased expression of TLR4 in colon adenomas and CRCs when compared to matched samples of normal colonic mucosa. In silico, expression of TLR4 was positively correlated with expression of COX-2, β-catenin, and EGFR. By IHC, 30% of CRCs have increased TLR4 expression compared with histologically normal epithelium obtained from the tumor margins. ImageJ analysis revealed that CRC's could be segregated based on fluorescent intensity into negative, intermediate, and strongly positive samples. Conclusions: TLR4 receptor expression defines a subset of patients with sporadic CRC and is associated with expression of key oncogenic pathways. Further study is needed to establish the relationship between TLR4 and CRC outcomes. No significant financial relationships to disclose.
Inactivation of O6 methylguanine DNA methyltransferase and nitrogen oxide in colorectal cancer.
Authors: Coudry R. A. , Assaf L. , Crespo R. , Santos E. M. 
Published: February 2017

461 Background: O6Methylguanine DNA methyltransferase (MGMT or AGT) activity plays an important role in colorectal carcinogenesis. Inactivation of MGMT can occur by promoter CpG island hypermetilation or through inhibition caused by nitrogen oxide (NO) and species derived from it. NO is synthesized from L-arginine by nitric oxide syntases (NOS) and plays a critical role in various biological processes and many disorders including cancer. Methods: We evaluated the expression of NOS1, NOS2, NOS3 and MGMT protein by immunohistochemistry. Plasma concentrations of nitrate/nitrite were analyzed by spectrophotometry. Methylation in the promoter region of MGMT was investigated by pyrosequencing. Results: Methylation of MGMT and loss of MGMT expression was detected in 68.8% and 32.6% of tumors, respectively. There was no significant association between MGMT methylation and protein expression with NOS1, NOS2 and NOS3 or nitrate/nitrite. Significant association was found between MGMT expression and methylation status (p < 0.001). NOS2 immunoreactivity was observed in the cytoplasm of the malignant cells (95.7%) and was positively associated with lymph node involvement (p = 0.046) and advanced stage (p = 0.048). Nitrite also showed association with lymph node involvement (p = 0.010). Conclusions: Loss of expression of MGMT protein is frequently caused by promoter hypermethylation at the MGMT gene and both are not associated with NOS enzymes or nitrate/nitrite. NOS2 overexpression and plasma concentration of nitrite may serve as a predictor for the prognosis of patients with CRC. No significant financial relationships to disclose.
Genetic profiles of hereditary nonpolyposis colorectal cancer.
Authors: Therklidsen C. , Jonsson G. , Bernstein I. , Nilbert M. 
Published: February 2017

469 Background: With the aim to identify genetic markers of hereditary nonpolyposis colorectal cancer (HNPCC), we applied tiling BAC array-based comparative genomic hybridization (aCGH) to 46 HNPCC-associated colorectal cancer. Methods: 32 k iling BAC arrays were used to generate high-density genomic profiles. Tumors were selected through a case-control design with half of the tumors derived from individuals with disease-predisposing mismatch repair gene mutations and the reminder from phenotypic HNPCC families without identified mutations. In addition, an equal number of sporadic tumors were used for comparison. Results: Tumors with disease-predisposing germline mutations showed frequent gains of chromosomes 1p (39%), 17 (43%), 19 (57%) and 22q (30%). HNPCC associated tumors without mutations did as a group have more complex alterations with the most frequent changes being gains of 20q (70%), 19 (35%), 17 (26%) and loss of 18 (39%). Gains of 1p and 20q and loss of chromosome 18 were identified as significant discriminators between HNPCC tumors with/without germline MMR gene mutations. Conclusions: The aCGH profiles of HNPCC-associated colorectal cancer suggest that specific gains and losses may be used to distinguish between tumors with/without germline mismatch repair gene mutations. No significant financial relationships to disclose.