bluuebunny/crossref_metadata_2025 huggingface dataset:
Preparation technique for cytologic study of EUS-guided FNA samples when onsite cytopathologist is absent: Cell block, smear, or both?
Authors: Nguyen N. Q. , Lim K. , Ruszkiewicz A.
Published: February 2017

176

Background: Smearing EUS-guided fine-needle aspiration (FNA) samples, with or without cell block, is most commonly performed for cytologic study in centers without onsite cytology. The diagnostic yield of such technique is often low (35%-50%) and may be related to loss of diagnostic material during the smearing process. Cell block technique captures not only aspirated cells but also small tissue fragments, improving diagnostic yield and enabling additional ancillary studies. This study aimed to compare the diagnostic yield of cell block alone against conventional smear (±cell block) technique in the EUS-guided FNA evaluation of pancreatic solid mass without onsite cytological assessment.

Methods: Data on the diagnostic yield of cell block alone technique performed over the last 12 months were prospectively collected and were compared to the yield of conventional smear ± cell block technique. A positive diagnostic yield was defined by the presence of adenocarcinoma and/or FNA findings consistent with the final surgical diagnosis.

Results: Data were available for 96 patients, of which 66 had cell block alone and 30 had smear +/-cell block preparation. The diagnostic yield of cell block alone was significantly higher that that of smear +/- cell block (53/66 vs 13/30; P=0.003). The addition of cell block after smearing did not improve the diagnostic yield (smear: 42% vs. both: 45%, p=0.88). Of patients who had smear +/-cell block, the diagnostic yield was significantly higher when the smearing was performed by cytopathologist as compared to that by nurse (10/16 vs. 3/14, p=0.02). There was a direct relationship between the rate of diagnostic yield and the number of FNA passes performed. The yield was highest when 4 or more FNA passes was performed (85%) as compared to that of 3 passes (63%, p=0.049) and 2 or less passes (38%, p<0.0001).

Conclusions: In the absence of onsite cytology services, the diagnostic yield from cell block alone was high (80%) and was superior to smear technique whether cell block was added to the evaluation of smears. In order to obtain adequate tissue material to maximise the diagnostic yield EUS-guided FNA of pancreatic mass, at least 4 FNA passes should be performed.

No significant financial relationships to disclose.
Detection of viable human circulating tumor cells using telomerase-specific GFP-expressing bioengineered adenovirus in patients with gastric cancer: A feasibility study.
Authors: Fujiwara T. , Uno F. , Hashimoto Y. , Shirakawa Y. , Nagasaka T. , Kagawa S. , Urata Y.
Published: February 2017

18

Background: The presence of circulating tumor cells (CTC) in the peripheral blood is associated with short survival and, therefore, the detection of CTC is clinically useful as prognostic factors of disease outcome and/or surrogate markers of treatment response. Recent technical advances in immunocytometric analysis and quantitative real-time PCR have made possible to detect a few CTC in the blood; however, there is no sensitive assay for detecting viable CTC. We developed a new approach to visually detect live CTC among millions of peripheral blood leukocytes using telomerase-specific replication-selective adenovirus expressing green fluorescent protein (GFP).

Methods: We constructed a GFP-expressing attenuated adenovirus, in which the telomerase promoter regulates viral replication (OBP-401, TelomeScan). The detection method for viable human CTC in the peripheral blood involves a three-step procedure including the lysis of red blood cells, the subsequent addition of OBP-401 to the cell pellets, and the automated scan under the fluorescent microscope. We analyzed fresh blood samples collected from 37 patients with histologically confirmed gastric cancer. We further assessed the CTC dynamics in patients who were undergoing chemotherapy or surgery to demonstrate the clinical potential of our approach for monitoring treatment responses.

Results: OBP-401 increased the signal-to-background ratio as a tumor-specific probe, because the fluorescent signal can be amplified only in viable human tumor cells by viral replication. Although the CTC level varied widely, ranging from 0 to 47 in 5-ml samples, 26 gastric cancer patients (70.3%) had more than one CTC; there was, however, no apparent relationship between CTC counts and TNM stages. Patients who had a recurrence of gastric cancer had decreased CTC counts after systemic chemotherapy. In the patients who underwent surgery, the CTC level dropped after complete resection.

Conclusions: This GFP-expressing virus-based method is simple and allows precise enumeration of CTC, which might be useful for monitoring the efficacy of local and systemic treatments.

[Table: see text]
Clinical significance of elevation of the serum IL-6 level in patients with advanced pancreatic cancer.
Authors: Mitsunaga S. , Ikeda M. , Nakachi K. , Ohno I. , Shimizu S. , Takahashi H. , Okuyama H. , Inagaki M. , Furuse J. , Ochiai A. ,   Chugai Pharmaceutical Co. Ltd.
Published: February 2017

181

Background: IL-6, one of the pro-inflammatory cytokines, is a recognized mediator of cachexia and cancer cell invasion. It has been reported that elevation of the serum IL-6 level may be associated with deterioration of the clinical condition and tumor progression in advanced pancreatic cancer (PC) patients. The aim of this study was to clarify the clinical features of increased serum IL-6 levels in patients with advanced PC receiving chemotherapy.

Methods: Patients with treatment-naïve unresectable PC and no obvious infectious conditions were eligible for this study. Serum levels of IL-6 were measured by an electrochemiluminescence assay. Symptoms were rated numerically from 0 to 10 using the Japanese version of the M. D. Anderson Symptom Inventory. Tumor volume was calculated as the sum of the long diameters of the tumors. The measurements were performed before chemotherapy and at one month after the start of chemotherapy.

Results: A total of 87 patients (male/female: 41/46; ECOG performance status: 0/1/2: 59/26/1; media age: 66 years) were enrolled; all patients were administered systemic chemotherapy (gemcitabine [GEM]/GEM+S-1/GEM+other/S-1: 52/11/9/15). The median serum level of IL-6 was 1.3 pg/mL before chemotherapy (at baseline) and 1.8 pg/mL at one-month after the start of chemotherapy. The median change of IL-6 from the baseline was +0.18 pg/mL. Patients with increase of the serum IL-6 level by more than 0.18 pg/mL were assigned to the elevated IL-6 group (n=42; median change in IL-6: +1.66 pg/mL). The elevated IL-6 group showed more sadness (p=0.019), numbness (p=0.008), and gain of body weight (p=0.016) at the baseline as compared to the non-IL-6-elevated group (n=42; median change in IL-6: -0.27 pg/mL). Comparison of the elevated and non-IL-6-elevated groups revealed a greater degree of increase in the tumor volume (p=0.015), deterioration of nausea (p=0.046) and vomiting (p=0.028), neutrophilia (p=0.004), and elevation of the serum C-reactive protein (p=0.011) in the elevated IL-6 group than in the non-IL-6-elevated group.

Conclusions: Elevation of the serum IL-6 level may be associated withsymptom deterioration, increase of the tumor mass, and inflammatory reaction in patients with advanced PC.

No significant financial relationships to disclose.
Power of fecal DNA methylation analysis: Does it lead to development of a noninvasive screening tool for pancreatic cancer?
Authors: Nagasaka T. , Goel A. , Taniguchi N. , Mori Y. , Sigeyasu K. , Yamada E. , Oka T. , Sun D. , Fujiwara T.
Published: February 2017

187

Background: The development of noninvasive screening tests is important to reduce mortality from gastrointestinal neoplasia, especially given that there is no noninvasive screening test for pancreatic cancer. We sought to develop such a test by analysis of DNA methylation from exfoliated cancer cells in feces.

Methods: We had reported a novel strategy that uses single-step modification of DNA with sodium bisulfite and fluorescence polymerase chain reaction methodology to measure aberrant methylation in fecal DNA. We increased number of biomarkers from 4 to 8 that were analyzed in 144 fecal samples obtained from a variety of patients, including colorectal tumors, hepatic tumors, and pancreatic tumors.

Results: The assay successfully identified one or more methylated markers in fecal DNA from patients with colorectal cancer, polyps, hepatocellular carcinoma, metastatic colorectal cancer at liver, and pancreatic cancer.

Conclusions: Methylation of 8 biomarkers in fecal DNA is associated with the presence of gastroenterological tumors relative to non-neoplastic conditions. Our fecal DNA methylation assay provides a possible means to noninvasively screen not only for colorectal tumors but also for pancreatic tumors.

No significant financial relationships to disclose.
Incidental diagnosis of pancreatic neuroendocrine tumors.
Authors: Cheema A. , Kvols L. K. , Strosberg J. R.
Published: February 2017

190

Background: Pancreatic neuroendocrine tumors are often discovered incidentally during radiologic or endoscopic examinations. The incidence of incidental detection is unknown. It is also unclear whether patients with incidentally discovered, asymptomatic tumors should be treated similarly to patients who present with tumor-related symptoms.

Methods: A database of 425 patients with pancreatic neuroendocrine tumors treated at the H. Lee Moffitt Cancer Center was developed. Patient charts were reviewed to assess whether their diagnosis was incidental or prompted by tumor-related symptoms such as pain, jaundice, or neuroendocrine hormone secretion. The frequency of “incidentalomas” was categorized by TNM stage (AJCC, 7th edition). Overall survival was stratified by “incidental” versus “symptomatic” diagnosis.

Results: Among 425 patients with histologically proven pancreatic neuroendocrine tumors, 112 patients (26%) had tumors that were discovered incidentally. The majority of stage I tumors (55%) were incidentally discovered. Among patients with stage IV tumors, 20% were detected incidentally (Table). Median survival of patients with incidentally discovered tumors was 103 months versus 84 months in patients who were symptomatic at diagnosis.

Conclusions: A sizeable fraction of patients with pancreatic neuroendocrine tumors are diagnosed incidentally during evaluations for other conditions or unrelated symptoms. The majority of patients with stage I tumors are incidentally diagnosed. The increased incidence of pancreatic neuroendocrine “incidentaloms” may be contributing to improving survival rates in this disease. This study highlights the necessity of developing guidelines for management of patients with incidentally discovered, early-stage tumors.

[Table: see text]

No significant financial relationships to disclose.
Myeloid-derived suppressor cells as a potential treatment target for pancreas adenocarcinoma.
Authors: Porembka M. R. , Mitchem J. B. , Goedegebuure P. S. , Linehan D.
Published: February 2017

194

Background: Myeloid-derived suppressor cells (MDSC) are a heterogeneous population of immunosuppressive cells that are upregulated in cancer. Little is known about the prevalence and importance of MDSC in pancreas adenocarcinoma (PA). Here, we quantify MDSC prevalence in patients with PA and assess the efficacy of MDSC depletion in a murine model of PA.

Methods: Peripheral blood and tumor samples were collected from patients with PA, analyzed for MDSC (CD15+11b+) by flow cytometry (FC) and compared to cancer-free controls (CFC). The suppressive capacity of MDSC and the effectiveness of MDSC depletion were assessed in C57BL/6 mice inoculated with Pan02, a murine PA, and treated with placebo or zoledronic acid (ZA), a potent aminobisphosphonate previously shown to target MDSC. Endpoints included tumor size, survival, and MDSC prevalence. Tumor cell infiltrate was analyzed by FC for MDSC (Gr1+CD11b+) and effector T cells; tumor cytokine levels were measured by Luminex assay.

Results: Patients with PA demonstrated increased circulating MDSC compared to CFC, which correlated with disease stage (metastatic PA: 68%±3.6% of CD45+ cells, resectable PA: 57%±3.5%, CFC: 37%±3.6%; p<0.0001). Normal pancreas tissue showed no MDSC infiltrate while PA avidly recruited CD11b+15+ cells to the primary tumor. Murine tumors similarly recruited MDSC that actively suppressed CD8+ T cells in vitro measured by CFSE dilution and accelerated tumor growth in vivo by adoptive transfer with Pan02 cells (p<0.001). Treatment with ZA impaired MDSC accumulation in the tumor (Placebo: 78%, ZA: 51%, p<0.05) resulting in delayed tumor growth rate (p<0.0001) and prolonged median survival (Placebo: 59 days, ZA: 73 days, p<0.05). MDSC blockade increased recruitment of T cells to the tumor (CD4: 4.4%±1.1% vs 12.2%±2.0%, p<0.05; CD8: 3.9%±1.3% vs 10.6%±2.2%, p<0.05) and a more robust type 1 response with increased levels of IFN-g (p<0.05) and decreased levels of IL-10 (p<0.05).

Conclusions: MDSC are an important mediator of tumor-induced immunosuppression in PA. Treatment with ZA effectively blocks MDSC accumulation improving anti-tumor response in animal studies. Efforts to block MDSC may represent a novel treatment strategy for PA.

[Table: see text]
Phase I trial of gene-mediated cytotoxic immunotherapy in combination with chemoradiation for locally advanced pancreatic cancer.
Authors: Bloomston M. , Marsh C. , Walker J. , Coyle W. , Marx H. , Tahiri S. , Moran Cruz C. , Aguilar L. K. , Aguilar-Cordova E. , Chung V. M.
Published: February 2017

195

Background: More than 80% of patients with pancreatic cancer present with locally advanced or metastatic disease and have a median survival of only 6 months. Immunotherapy approaches may improve outcomes. Gene Mediated Cytotoxic Immunotherapy (GMCI) is an approach that generates a systemic anti-tumor vaccine effect through intra-tumoral delivery of an adenoviral vector expressing the HSV-thymidine kinase gene (AdV-tk) followed by anti-herpetic prodrug and synergy with chemoradiation. The mechanisms of action involve tumor cytotoxicity, activation of antigen presenting cells and stimulation of systemic anti-tumor T-cell immunity. Safety with potential efficacy has been demonstrated in multiple clinical studies. This is the first application of GMCI in pancreatic cancer.

Methods: This study evaluated 4 dose levels of AdV-tk (3x1010 to 1x1012 vector particles) injected into locally advanced tumors via EUS or CT-guidance before and during week 3 of standard 5-FU-chemoradiation. Valacyclovir (Valtrex, GSK) prodrug was given for 14 days after each of 2 AdV-tk injections.

Results: The study completed accrual with 13 patients enrolled and 12 completing therapy with 3 at each of the 4 dose levels. One patient refused further participation during course 1 after recovering from azotemia. Median age was 64 years (range 55-81) and median baseline CA19-9 was 1634 U/ml. No dose limiting toxicities and no injection related complications occurred. Possibly related grade 3-4 toxicities, all of which were transient, included dehydration, azotemia and worsening elevation of bilirubin and AST. Kaplan-Meier estimated median survival is 12.2 months with 6 patients still alive at 8-20 months. Two patients achieved a partial response by RECIST criteria. One occurred in week 6 despite discontinuing 5-FU/radiation during week 1. The other had gradual decrease of a 7 cm tumor over 11 months. Serum CA19-9 levels decreased in 8/8 evaluable patients by 32-91% at 3 months after treatment initiation.

Conclusions: AdV-tk can be safely injected into pancreatic tumors and combined with standard chemoradiation. Early results are highly encouraging and justify further evaluation in a phase II study.

[Table: see text]
Molecular pathologic phenotypes and outcome in adenocarcinoma of the ampulla of Vater.
Authors: Chang D. K. , Johns A. , Scarlett C. J. , Pajic M. , Chou A. , Toon C. , Samra J. S. , Sutherland R. , Gill A. J. , Biankin A. V. ,   New South Wales Pancreatic Cancer Network
Published: February 2017

196

Background: Individuals with adenocarcinoma of the ampulla of Vater demonstrate a broad range of outcomes, as they may arise from any of the three epithelia that converge at that location. This variation in outcomes poses specific challenges in clinical decision making concerning the aggressiveness of treatment, and the appropriateness and type of adjuvant therapy.

Methods: We assessed the relationship between molecular pathologic phenotypes defined using a combination of histopathology and protein expression (CDX2 [caudal-type homeodomain transcription factor 2] and MUC1), and outcome in 72 patients who underwent operative resection for adenocarcinoma of the ampulla of Vater.

Results: Patients with a pancreaticobiliary phenotype (CDX2 negative, MUC1 positive) compared to an intestinal phenotype carcinoma segregated into an independently poor prognostic group (HR = 3.40, 95% CI: 1.71 – 6.76, p = 0.0005). Stratification using lymph node (LN) status (the only other independent poor prognostic factor) (HR = 3.19, 95% CI: 1.54 – 6.58, p = 0.0017) defined three clinically relevant phenotypes: 1) those with a non-pancreaticobiliary (intestinal) phenotype without LN metastases who had an excellent outcome (5-year survival 88.4%, median survival 172.8 months); 2) those that had a pancreaticobiliary phenotype and lymph node metastases that had a poor outcome (5-year survival 20.0%, median survival 7.4 months); and 3) the remainder (intestinal/LN positive or pancreaticobiliary/LN negative) who had an intermediate outcome (5-year survival 46.9%, median survival 57.0 months).

Conclusions: A combination of histopathologic and molecular criteria identified three distinct clinically relevant phenotypes of adenocarcinoma of the ampulla of Vater that have significantly different outcomes. This distinction, which requires validation, may be used to refine current therapeutic strategies by guiding aggressiveness of surgery and selection for adjuvant therapy, which would improve overall outcomes.

No significant financial relationships to disclose.
Effect of the ultraconserved noncoding RNA uc.338 on cellular growth of hepatocarcinoma.
Authors: Braconi C. , Kogure T. , Valeri N. , Gasparini P. , Huang N. , Nuovo G. , Terracciano L. , Croce C. M. , Patel T.
Published: February 2017

202

Background: The role of non-protein coding (nc)RNAs in cancer is unknown but emerging evidence suggests that deregulated expression of ncRNA may contribute to cancer pathogenesis. We sought to examine the role of ultraconserved ncRNA (ucRNA) that are 100% conserved across the human, rat and mouse genomes in hepatocellular cancers (HCC).

Methods: Whole genome ucRNA expression profiling was performed using a custom microarray, and verified by real time PCR in cell lines and by in situ hybridization in a tissue microarray comprising of 221 human HCC, 72 non cirrhotic (NC) and 97 cirrhotic (C) liver tissues. ucRNA expression was manipulated with siRNA or plasmid-over-expressing ucRNA, and the effects on anchorage-dependent and independent growth, and cell cycle assessed using cell viability, soft agar assays and flow cytometry. Gene ontology analysis was performed by evaluating uc338-dependent changes on mRNA expression using Affymetrix chips.

Results: 56 ucRNAs were aberrantly expressed with 33 increased and 23 decreased in HepG2 cells compared to non-malignant hepatocytes. The greatest change was observed with uc.338 (6.9-fold increase). uc.338 expression was significantly increased in several HCC cell lines. uc.338 expression was detected in 170 cases (77%) of HCC, with 62% of these showing a moderate to strong expression. Compared to non- malignant adjacent tissue, uc.338 expression was increased in 97/156 of HCC. The mean % of cells expressing uc.338 was 4% in NC liver, 15% in C and 24% in HCC. Inhibition of uc.338 reduced anchorage dependent and independent growth, and cell cycle progression in both human and murine malignant hepatocytes. Gene annotation enrichment analysis of mRNAs that were altered by inhibition of uc.338 expression identified the top over-represented GenMAPP pathways as: cell cycle, mRNA processing, RNA transcription, G1 to S cell cycle. Moreover, enforced expression of uc.338 increased cell growth in nonmalignant hepatocytes.

Conclusions: These data showing that uc.338 is selectively overexpressed in HCC and promotes cell growth provides new insights into the role of RNA genes in HCC.

No significant financial relationships to disclose.
The influence of CD44v6, TGF-α, COX-2, MMP-7, and MMP-9 on clinical evolution of patients with gastric cancer.
Authors: Fanelli M. F. , Chinen L. T. , Begnami M. D. , Costa W. L. , Fregnami J. H. , Soares F. A. , Montagnini A. L.
Published: February 2017

21

Background: Gastric cancer (GC) is the fourth most common tumor worldwide. As palliative chemotherapy is a relevant approach for patients with distant metastasis, determination of prognostic parameters that are predictive of survival may allow better identification of patients who may benefit from current chemotherapy regimens. The process of GC proliferation and metastasis occurs via epithelial-mesenchymal transition (EMT), which consists on gain of motility and invasiveness by epithelial cells via the disassembly of cell-to-cell contacts and significant cytoskeletal reorganization. Molecules involved in each step have been identified, including cell-adhesion molecules, growth factors and matrix degradation enzymes. Objectives: To verify the correlation between tumor progression and the expression of some proteins (CD44v6, TGF-α, COX-2 MMP-7, MMP-9) involved on EMT, by these relation with patient overall survival (OS) and disease stage.

Methods: We retrospectively examined the medical records and histopathological biopsy results of 137 patients with GC (stages I to IV), treated at the Department of Medical Oncology at Hospital do Cancer A.C. Camargo between 1998 and 2006. Included patients underwent total or subtotal gastrectomy at our hospital. Expression of proteins related to EMT was evaluated by Tissue Microarray.

Results: By the analysis of protein expression according to deep of gastric wall invasion (T), no protein showed to be important. On different levels of lymphonodal commitment (N0, N1, N2, N3) and in relation to presence (M1) or absence (M0) of distant metastasis, only COX-2 expression showed to be determinant with major expression on N2 and N3 and M1 status (p = 0.017 and p = 0.011, respectively). COX-2 was also relevant for clinical stage, where its expression was higher on poor stages (p = 0.011). On 5-year OS, TGF-α (p = 0.017), COX-2 (p < 0.001), MMP-7 (p = 0.020) and MMP-9 (p = 0.015) expressions revealed to be determinant, where the poor expression of these proteins were linked to better OS.

Conclusions: This is the first study to correlate proteins involved on EMT with OS. We were able to show that COX-2, TGF-α, MMP-7 and MMP-9 expression on GC were related to poor patient OS.

No significant financial relationships to disclose.
The optimal tumor response assessment in patients with HCC after treatment of TACE.
Authors: Jung E. , Kim J. , Yoon E. , Lee H. , Lee Y. , Yeon J. , Byun K.
Published: February 2017

215

Background: WHO or RECIST has been considered as suboptimal methods for tumor response assessment in HCC, especially receiving locoregional therapies. Recently, new modified methods having concept of viable tumor has been proposed. Herein we compared the efficacy to assess response and to predict prognosis between WHO, RECIST, EASL, mRECIST in HCC patients receiving TACE.

Methods: We retrospectively reviewed 141 HCC patients who were newly diagnosed and received TACE between August 2005 and November 2006. Among them, 98 patients who were available for assessing treatment response were analyzed at 1 month after 2nd TACE. We evaluated intermethod aggreement between methods and assessed the efficacy predicting time to progression (TTP) and overall survival (OS).

Results: The results of tumor responses (number of CR/ PR/ SD/ PD) by WHO/ RECIST/ EASL/ mRECIST were 1/ 14/ 74/ 9, 1/ 12/ 77/ 8, 34/ 34/ 25/ 5, 34/ 28/ 31/ 5, respectively. The κvalue of intermethod agreement for EASL vs WHO, EASL vs RECIST, mRECIST vs WHO, and mRECIST vs RECIST was 0.102, 0.088, 0.112 and 0.122, respectively. The κvalue for WHO vs RECIST and EASL vs mRECIST was 0.759 and 0.883 (p=0.00). Hazard ratios for TTP in responders compared with nonresponders were 0.313 (95% CI; 0.19-0.52, p=0.00) for EASL and 0.335 (95% CI; 0.21-0.54, p=0.00) for mRECIST. Hazard ratios for survival in responders compared with nonresponders were 0.208 (95% CI; 0.12-0.37, p=0.00) for EASL and 0.269 (95% CI; 0.15-0.48, p=0.00) for mRECIST. In the univariate analysis, tumor size, infiltrative type, PVT, EASL responder, and mRECIST responder were significant risk factors for survival. Multivariate analysis showed EASL responder (HR 0.165, 95% CI 0.056-0.486, p=0.001) and infiltrative type (HR 4.504, 95% CI 1.579-12.848, p=0.005) were independent predictable factors for survival.

Conclusions: The recent proposed methods showed different characteristics from preexisting methods in assessing treatment response and furthermore, responder by EASL method was independent predictor for survival in HCC patients receiving TACE. EASL method should be applied as response assessment method for HCC patients receiving TACE.

No significant financial relationships to disclose.
Effect of nicotine on chemoresistant phenotype as mediated through Src-dependent Id1 expression in pancreatic adenocarcinoma cells.
Authors: Trevino J. G. , Pillai S. R. , Chellappan S. P.
Published: February 2017

216

Background: The signaling pathways contributing to DNA-binding protein inhibitor Id1 expression and chemoresistance in pancreatic cancer remain unknown. Id1 plays a role in pancreatic tumor progression with tumor-promoting effects of nicotine regulating protein tyrosine kinase Src activation and Id1 expression, both associated with chemoresistance in other systems. We hypothesize Id1 expression regulates chemoresistance in pancreatic cancer through a nicotine-promoting Src-dependent pathway.

Methods: We probed pancreatic cancer cell lines (L3.6pl, PANC-1, Mia-PaCa-2) for innate gemcitabine chemoresistance with cell viability MTT assay and western blot analysis of PARP cleavage programmed cell death. Gemcitabine-sensitive cells were exposed to rising gemcitabine concentrations to establish a resistant subtype, L3.6plGemRes. Protein analysis and mRNA expression were determined by western blot analysis and RT-PCR respectively. Induction of Src phosphorylation or Id1 expression was performed with nicotine (1 μM).

Results: Inhibition of c-Src expression was performed with short-interfering RNA (siRNA). Nicotine-induced Src phosphorylation and Id1 expression. Inhibition of Src by siRNA resulted in decreased nicotine-induced Id1 expression. Inhibition of Src and Id1 expression by siRNA in innate or established gemcitabine resistant pancreatic cancer cells resulted in gemcitabine sensitization. To determine if nicotine contributes to gemcitabine chemoresistance, we exposed gemcitabine-sensitive cells to nicotine with subsequent exposure to gemcitabine IC50, 250 ng/ml, and cell viability assays confirmed a 2-fold increase in cell prolilferation and a 4.5-fold reduction in apoptosis. Further, nicotine induced phosphorylation of key signaling enzymes involved in proliferation and apoptosis, Erk1/2 and Akt respectively.

Conclusions: In summary, we demonstrate that Id1, through a nicotine-promoting Src-dependent pathway, is necessary for establishment of a chemoresistant phenotype in pancreatic cancer cells. Understanding the signaling pathways involved in pancreatic tumor chemoresistance will lead to therapies resulting in improved tumor responses.

No significant financial relationships to disclose.
VEGF receptor 1 and heparanase expression as prognostic factors in cholangiocellular carcinoma.
Authors: Goekkurt E. , Stoehlmacher J. , Aust D. E. , Danenberg P. V. , Danenberg K. D. , Lehmann S. , Mogck U. , Ehninger G. , Hoffmann A. C.
Published: February 2017

226

Background: Cholangiocellular carcinoma (CCC) remains to be a tumor with very few treatment choices and limited prognosis. In this study, we sought to determine the prognostic role of VEGF receptor 1 (VEGFR1), heparanase (HPSE) and EGF receptor (EGFR) gene expression in patients with resected CCC.

Methods: 47 formallin-fixed paraffin embedded FFPE tumor samples from patients with resected CCC were analyzed. FFPE tissues were dissected using laser-captured microdissection and analyzed for VEGFR1, HPSE and EGFR mRNA expression using a quantitative real-time RT-PCR method. Gene expression values (relative mRNA levels) are expressed as ratios between the target gene and internal reference genes (beta-actin, b2mg, rplp2, sdha).

Results: VEGFR1 and HPSE expression levels were significantly associated with overall survival (OS). Patients expressing low levels of VEGFR1 displayed a median OS of 23.2 months compared to 5.3 months in patients with high VEGFR1 expression (p=0.006, logrank test). Patients with low HPSE expression had a median OS of 19.8 months compared to 10.0 months in patients with high HPSE expression (p=0.02, logrank test). EGFR and PDGFRα expression were significantly associated with HPSE and VEGFR1 expression (p=0.03 and p<0.0001, respectively).

Conclusions: These results suggest that VEGFR1 and HPSE gene expression levels may play a role as prognostic factors in patients with CCC. Further studies are warranted to study these associations.

[Table: see text]
The impact of bevacizumab on liver regeneration following hepatectomy in rats.
Authors: Mori H. , Shimada M. , Utsunomiya T. , Imura S. , Morine Y. , Ikemoto T. , Hanaoka J. , Iwahashi S. , Saito Y. , Yamada S.
Published: February 2017

228

Background: In our clinical data, we had the interesting findings that the pathologic sinusoidal obstruction syndrome and serum hyaluronic acid after neoadjuvant oxaliplatin-associated chemotherapy for liver metastatic colorectal cancer with bevacitumab (Bev) was ameliorated compared to those without Bev. The purpose of this study was to investigate the influence of bevacizumab administration on regenerating liver in rat 70% and 90% hepatectomy (Hx) model as a surrogate model of human massive hepatectomy for liver metastatic colorectal cancer.

Methods: Male Wister rats weighing 180-230g were divided into the following four groups: 70%Hx, 70%Hx + Bev, 90%Hx and 90%Hx+Bev group. The rats were pretreated with intraperitoneal administration of bevacizumab (5mg/kg) 7 days before hepatectomy. The remnant liver and blood samples were taken one day after hepatectomy, and the following parameters were evaluated: blood analysis (AST, ALT, LDH, T- Bil, and hyaluronic acid), liver weight to body weight (Lw/Bw) ratio, and postoperative survival rate for three days.

Results: In the 70%Hx model, there was no significant difference between the 70%Hx group and 70%Hx + Bev group in blood analysis one day after hepatectomy; AST (1928 vs. 923 IU/L), ALT (1282 vs. 670 IU/L), T-Bil (0.17 vs. 0.19 mg/dl), LDH (3822 vs. 2967 U/L) and hyaluronic acid (995.7 vs. 1026.6 ng/ml) and in Lw/Bw ratio (1.78 vs. 1.84). In 90%Hx model, AST and ALT of blood analysis in 90%Hx+Bev group significantly decreased compared to those in 90%Hx group; AST (3428 vs. 4995 IU/L, P<0.05), ALT (1471 vs. 2287 IU/L, P<0.05), T-Bil (1.94 vs. 2.57 mg/dl), LDH (3229 vs. 5351 U/L) and hyaluronic acid (3865.7 vs. 7862.2 ng/ml). The Lw/Bw ratio in 90%Hx+Bev group significantly increased compared to that in 90%Hx group (0.80 vs. 0.67, P<0.05). The postoperative survival rate in 90%Hx+Bev group showed tendency to improve compared to that in 90%Hx group (75% vs. 50%, P=0.19).

Conclusions: The administration of bevacizumab seven days before hepatectomy did not significantly affect the liver functions and liver regeneration rate. These findings suggest that hepatectomy might be safe and feasible after the use of bevacizumab.

No significant financial relationships to disclose.
Molecular mechanisms underlying the synergistic interaction of the novel anticancer drug celandine with gemcitabine in preclinical models of pancreatic cancer.
Authors: Funel N. , Giovannetti E. , Del Chiaro M. , Pollina L. , Mosca F. , Peters G. , Campani D. , Boggi U.
Published: February 2017

233

Background: Celandine (Ukrain) increased median survival in combination with gemcitabine compared to gemcitabine alone (10.4 vs. 5.2 months; p<0.001) in unresectable PDAC (Gansauge et al.; Langenbecks Arch Surg 2002). There is compelling evidence that gene transcripts of determinants of gemcitabine activity could be used to tailor PDAC chemotherapy (Giovannetti et al., Cancer Res 2006).

Methods: In vitro studies were performed in two ATCC cell lines (PL45 and Mia PaCa-2) and two primary cell cultures obtained from PDAC patients who underwent surgical resections (PPTCC78 and PPTCC109). Cells were treated with celandine at IC50concentration levelsfor 48 h. The total RNA extraction was performed with TRIzol protocol. All the amplifications were carried out with normalization of gene expression against the glyceraldehyde 3-phosphate dehydrogenase (GAPDH) housekeeping control gene, and the quantitation of gene expression was performed using the direct ratio, the standard curve method and the ΔΔCT calculation, in which the amount of target, normalized to the endogenous control and relative to the calibrator (untreated control cells) was calculated as 2(-ΔΔCt).

Results: Celandine positively modulates the expression of hENT1 mRNA in all PDAC cell cultures treated with IC50 (p<0.001). The 2(-ΔΔCt) analysis revealed a mean increase of 2.8-fold (p=0.001) with respect to untreated control cells. In PL45 and Mia PaCa-2 cells celandine positively affects mRNA expression of dCK gene as well.

Conclusions: To date a few options based on gemcitabine are available for treatment of PDAC. Most gemcitabine-based chemotherapy regimens resulted in a very limited disease control, and studies attempting to widen the therapeutic armamentarium against this disease are warranted. Based on the previous clinical data the celandine-gemcitabine combination appears a promising regimen and the results of the present study provide the experimental basis for the further clinical testing of the celandine-gemcitabine schedule in PDAC patients.

No significant financial relationships to disclose.
Phase II trial of neoadjuvant full-dose gemcitabine, oxaliplatin, and radiation (RT) in patients with resectable (R) or borderline resectable (BR) pancreatic cancer.
Authors: Kim E. J. , Ben-Josef E. , Griffith K. A. , Herman J. M. , Wolfgang C. L. , Bekaii-Saab T. S. , Bloomston M. , Dawson L. A. , Moore M. J. , Zalupski M. M.
Published: February 2017

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Background: We previously developed a regimen of full dose gemcitabine (gem), oxaliplatin (ox) and RT to maximize systemic and loco-regional disease control in pancreas cancer (JCO 25:4587, 2007). A multi-institution phase II trial was conducted to test efficacy of this regimen as neoadjuvant therapy in R/BR disease.

Methods: Eligibility criteria included path confirmation, no metastasis, R/BR lesion, PS ≤2, and adequate organ function. Treatment consisted of 2, 28 day cycles of gem (1g/m2 over 30 min D1, 8, 15) and ox (85mg/m2 D1, 15) with RT during cycle 1 (30Gy in 2Gy fractions). Pts were then re-evaluated for surgery. Resected pts received 2 additional cycles of chemotherapy.

Results: 68 evaluable pts were treated at 4 centers in 2007-2010. Median age was 64 (42-83), 32 men, PS 0:1:2 in 40:26:2. Median tumor size 3.2 cm (1.4–7.8), lesion in head 49, body 9, tail 10, R in 24 and BR in 44. 66 pts (97%) completed cycle 1/RT and 61 (90%) cycle 2. Therapy related adverse events ≥ grade 3 in cycles1/2 included ANC (32%), plts (24%), GI (16%), biliary/cholangitis (15%). Best response in primary was partial (10%) or stable (81%). 20 pts not operated on protocol: 8 progression (4 local, 4 distant), 8 judged not resectable, 3 toxicity, 1 early death. Of 48 laparotomies,10 not resected due to vascular involvement (6) or M1 disease (4). Resection completed in 15 of 18 R pts (13 R0, 2 R1) and 23 of 30 BR pts (19 R0, 2 R1, 2 R2). 26 pts received post-op therapy. With median fu 11.3 mos (0.7-35), 42 pts are alive, 20 pts are NED. Median survival (OS) for all pts is 21.2 mos (95%CI 13.3-not defined [ND]), resected 31.1 mos (95%CI 13.7-ND), unresected 16.0 mos (95%CI 5.8-ND). Time to treatment failure (death, progression, toxicity, no resection) and OS in R pts are 9.1 mos (95%CI 2.4-23.8) and 31.1 mos (95%CI 9.8-ND) and in BR pts 5.5 mos (95%CI 2.4-11.8) and 18.0 mos (95%CI 13.3-ND). Correlation of pathologic response and outcome is ongoing.

Conclusions: Neoadjuvant therapy with full dose gem, ox and RT was possible in a large proportion of pts with localized pancreas cancer and resulted in a high percentage of R0 resections. Results are particularly encouraging given a majority of pts with BR disease.

[Table: see text]
Efficacy of lanreotide 30 mg as symptomatic treatment in patients with inoperable bowel obstruction due to peritoneal carcinomatosis: A randomized double-blind, placebo-controlled study.
Authors: Mariani P. , Blumberg J. , Chauvenet L.
Published: February 2017

242

Background: Somatostatin analogues in previous open label studies improved symptoms due to bowel obstruction in patients with peritoneal carcinomatosis. This study assessed the efficacy and safety of lanreotide microparticles 30 mg as treatment of clinical symptoms in inoperable patients.

Methods: Eighty cancer patients with inoperable digestive obstruction of malignant origin (mean age 62.3 years, 82.5% female) were randomized to receive one intramuscular injection either of lanreotide microparticles 30 mg (N=43) or placebo (N=37). The primary location of cancer was mostly genital (ovary 37.5%, uterus 13.8%) or digestive (colon 16.3%, stomach 11.3%, pancreas 6.3%). Most patients were fed by central parenteral route (80%) and were severely impaired (63.8% ECOG 3 or 4). The primary endpoint was the response rate at day 7 (responders defined as patients with ≤ 1 vomiting episode per day or no vomiting recurrence after removal of the NGT, for at least 3 consecutive days) assessed in a diary card in the intention to treat (ITT) population. Per protocol (PP) population mainly excludes concomitant treatment and inclusion criteria deviations defined during a blinded data review meeting.

Results: In the ITT analysis, 41.9% of lanreotide 30mg treated patients were responders versus 29.7% for placebo. This difference was not statistically significant (odds ratio (OR) = 1.75 (95% CI [0.68; 4.49], p = 0.24, logistic regression). The predefinedPP (n= 49) sub group analysis did show a statistically and clinically significantly higher response rate in the lanreotide group (OR 3.60 (95% CI [1.03, 12.62], p = 0.045). There was no significant difference between the 2 treatment groups in secondary efficacy endpoints assessment but there were significant differences in favor of lanreotide in well-being using a visual analogue scale at D3 (p = 0.04), D6 (p = 0.04) and D7 (p = 0.01). No drug-related serious adverse event (SAE) or unexpected event was reported.

Conclusions: Lanreotide 30 mg may decrease clinical symptoms and improve well-being in patients with inoperable bowel obstruction due to peritoneal carcinomatosis.

[Table: see text]
Phase II study of sunitinib malate following hepatic artery embolization for metastatic neuroendocrine tumors.
Authors: Strosberg J. R. , Cheema A. , Campos T. , Valone T. , Kvols L. K.
Published: February 2017

244

Background: Neuroendocrine tumors (NETs) frequently metastasize to the liver. Hepatic arterial embolization is an important therapeutic modality in treating patients with liver-predominant metastases. NETs are highly vascular and are known to express both VEGF and VEGFR. We hypothesize that administration of sunitinib, a VEGFR inhibitor, following hepatic artery embolization will delay tumor revascularization and extend progression-free survival.

Methods: Patients with metastatic neuroendocrine tumors to the liver underwent a series of selective arterial embolizations followed by sunitinib (one week after each embolization, and continued until disease progression or up to a maximum of 8 cycles). Radiographic response rates were assessed by RECIST criteria. PFS and OS were calculated using Kaplan-Meier methodology.

Results: 39 patients with metastatic NETs were enrolled. Primary tumor sites included the small intestine (26), pancreas (10), rectum (2), and lung (1). The initial starting dose of sunitinib was 50 mg; however, all five patients enrolled at this dose required dose reductions, and the starting dose was subsequently lowered to 37.5 mg. The majority of patients required further dose reductions to 25 mg. Twenty-eight patients (72%) experienced a partial radiographic response (PR), eight patients (20%) had stable disease, and three patients (8%) had progressive disease as their best response. Median PFS was 18 months and the rate of 1-year PFS was 72%. The rates of overall survival (OS) at 1 year and 2 years were 94% and 78%. Serum VEGF levels increased by an average of 51 pg/ml (34%) after embolization.

Conclusions: Hepatic artery embolization is a highly active treatment option for patients with metastatic neuroendocrine tumors to the liver. Embolization stimulates release of VEGF into the circulation. Sunitinib can be administered following hepatic artery embolization. The high rates of progression-free survival and overall survival associated with this sequence of therapies are encouraging.

[Table: see text]
The outcome of definitive or salvage radiotherapy for locally advanced hepatocellular carcinoma: A retrospective cohort study of 822 patients from a single institution.
Authors: Seong J.
Published: February 2017

248

Background: With technological development and awareness of efficacy, radiotherapy (RT) is more frequently adopted in management of hepatocellular carcinoma (HCC). To assess the efficacy of RT, we conducted this retrospective cohort study from a single institute.

Methods: The analysis involved 822 patients who underwent definitive or salvage RT for locally advanced HCC from January 1997 to August 2009. Two-dimensional RT, 3-dimensional conformal RT (3D-CRT), and intensity-modulated RT including tomotherapy (IMRT) were carried out for 186 (22.6%), 579 (70.4%), and 57 (6.9%) patients, respectively. In a majority of patients, RT was done either concurrently with intra-arterial 5-FU (500 mg/m2) chemotherapy (CCRT group; 326 patients, 39.7%), or following transarterial chemoembolization (TACE+RT group; 244 patients, 29.7%). Total radiation dose was 30 to 64.8 Gy (median dose 45 Gy) in 1.8-2 Gy fraction.

Results: The median age of total 822 patients was 55 and 84.3% was male. The 2, 3, and 4-year overall survival rates of total patients were 21.2, 13.1, and 10.1%, respectively. In 2-year survival, the patients with Child-Pugh class A did better than B or C (23% vs 10.9%, p<0.001), without portal vein thrombosis (PVT) did better than with PVT (26.8% vs 14.3%, p<0.001), and without lymph node metastasis (LNM) did better than with LNM (22.9% vs 11.7%, p<0.001). Patients with total dose higher than 45 Gy did better than those with less than 45 Gy (30.1% vs. 15.6%, p<0.001). The best outcome was shown in patients received higher than 45 Gy using 3D-CRT or IMRT, with 2-year overall survival rates of 25.9% (CCRT) and 41.3% (TACE+RT).

Conclusions: This study showed a substantial effect of RT in locally advanced HCC. Further analysis will be continued to provide the best option of radiotherapy for locally advanced HCC.

No significant financial relationships to disclose.
Prevalence of HER2/neu overexpression/amplification in a Hispanic population with gastric adenocarcinoma.
Authors: Rodarte Shade M. , Flores Gutierrez J. , Garcia Labastida L. , Villela L. , Barbosa Quintana A. , Paredes Garcia A. , Gamboa O. , Guzman Huerta E.
Published: February 2017

25

Background: In Europe, Asia, and North America, studies have identified that approximately 20% of patients with gastric adenocarcinoma exhibit overexpression of HER2/neu. This group of patients has been shown to benefit from combination therapy with the anti-HER2/neu monoclonal antibody trastuzumab. These findings have not been studied in Hispanics who may possess a distinct gastric cancer genotype. The aim of this study is to determine the rate of HER2/neu positivity in a completely Hispanic population with gastric adenocarcinoma and to analyze the clinical factors associated with HER2/neu.

Methods: We conducted a retrospective study in three different hospitals in Monterrey, Mexico. The study population consisted of Hispanic patients with gastric adenocarcinoma who have had a gastric resection and for whom there were adequate amounts of tissue available in paraffin blocks. Tissue was tested for HER2/neu using both Immunohistochemistry (IHC) and the fluorescent in situ hybridization (FISH) techniques.

Results: From 2000 to 2010, we initially evaluated 27 gastric cancer tumor samples. Sex distribution was 15 males and 12 females with a mean age of 57±15 (SD) years. Anatomic distribution of tumor was 4% in the cardias, 40% in the body and 56% in the antrum. Lauren's histological type distribution was 52% diffuse and 48% intestinal. Fifty-three percent of the tumors were high grade and nodal positivity was present in 69%. Overexpression of Her-2/neu (IHC ++ and +++) was found in five (17.4%) of the samples, being moderate (++) in two (7.4%) and strong (+++) in three (11%) tumor samples. FISH amplification for the HER2/neu gene was found in two (7.4%) tumors and was only seen in samples with a strong (+++) IHC result. There was no association identified between HER2/neu status with age, gender, degree of differentiation, T stage, or nodal status.

Conclusions: In a homogeneous Hispanic population, 17.4% of gastric cancer patients were found to overexpress HER2/neu by IHC. FISH did not identify any additional HER2/neu positive tumors. It is important to consider IHC testing for HER2/neu in Hispanic gastric cancer patients as they could benefit from a chemotherapy regimen containing trastuzumab.

No significant financial relationships to disclose.