M1233 Expression of Growth Factor Receptor and Potential Treatment in Cholangiocarcinoma (CC) Cell Lines Ling Xu, Theresa Pesch, Silvia Kellermeier, Werner Falk, Jürgen Schölmerich, Michael Fried, Frank Klebl, Gerhard Rogler Introduction: CC is a rare carcinoma of the biliary epithelium. Standard chemotherapy has little effect on CC. Growth factors and their receptors play an important role in tumor growth and metastasis. Neutralizing antibodies targeting growth factors and their receptors have been shown to be therapeutically effective. We therefore determined the expression of 7 growth factors in 4 human CC cell lines. The effect of monoclonal EGFR antibody on two human CC cell lines was tested. Methods: mRNA expression was studied for EGFR, VEGFR1, VEGFR2, VEGFR3, IGF1R, IGF2R and HGFR in the four human CC cell lines TFK-1, EGI-1, OZ and HuH28. Protein expression of these molecules was investigated by Western blot and immunohistochemistry. The effects of 0.1, 1.0, 10, 100 and 1000 mg/ml monoclonal anti-EGFR antibody (cetuximab, ErbituxÒ, Merck) were tested In Vitro in the two cell lines TFK-1 and EGI-1. Cell growth was analyzed by cell counting, cell metabolism by XTT assay and apoptosis by FACS. Results: mRNA and protein expression of all seven growth factor receptors was found in all four human CC cell lines by PCR, Western blot and immunohistochemistry. Cetuximab significantly inhibited cell growth of TFK-1 in a dose dependent manner (p < 0.05) but had no influence on EGI-1 cells. The metabolic activity of TFK-1 cells treated with cetuximab was increased dose dependently, however, decreased significantly over time. For EGI-1 cells metabolic activity remained largely unchanged. TFK-1 but not EGI-1 showed increased rates of apoptosis (p < 0.05). Results: Cetuximab has different major effects on cell growth, cell metabolism and apoptosis on human CC cell lines TFK-1 and EGI-1. Cetuximab inhibits TFK-1 cell growth and increased apoptosis. These effects are not related to the expression of growth factor receptors. M1234 Randomized Prospective Study with a Intravenous Anesthesia (Propofol) for Radiofrequency Thermal Ablation (RFA) in Hepatic Neoplasms Shinpei Sato, Akishige Yamada, Tatsuya Minami, Shintaro Mikami, Akemi Takada, Tenyu Motoyama, Satoru Yotsuya, Teruyuki Tanaka, Shintaro Kondo, Nobuo Toda OBJECTIVES : Radiofrequency thermal ablation(RFA) is a minimally invasive treatment widely performed for the treatment of liver neoplasms in Japan ; however, it is generally performed under local analgesia and patients, as a rule, complaint severe pain during the procedure. Propofol has pharmacokinetic characteristic suitable for use as a total intravenous anesthetic agent. But there are no studies concerning the feasibility and safety of propofol sedation in RFA-treated patients. The aim of this study was to elucidate whether the short- acting anesthetic propofol offers certain potential advantages compared with conventional sedation during RFA. METHODS : Sixty patients with liver neoplasms, randomly assigned receive midazolam plus pentazocine (n = 32) or propofol alone (n = 28) for sedation during RFA. Vital signs were continuously monitored and procedure-related parameters such as recovery time, patients' cooperation, analgesia, tolerance of the procedure, anoxemia, pres- ence of hypertention or hypotention, presence of postoperative complication, dosage of sedative agent and pain degree, stress of practitioner were assessed. The degree of a pain during procedure was noted by Numeric Rating Scale(NRS) on a visual analogue scale ranging from 0 to 10 points. Patients was administrated any dose of sedative agent considered appropriate for the patient's age and size. RESULTS : Propofol provided a significantly better patient cooperation and analgesia than midazolam/ pentazocine (p< 0.01), also was significantly milder in the degree of a pain during procedure. Mean recovery time was significantly shorter in the propofol group (45 ±17 min vs 120 ± 48 min) (p< 0.01).Also blood pressure significantly elevated in the midazolam/ pentazocine group than propofol group. Frequency of anoxemia was higher in the midazolam/ pentazocine group (0% vs 14%). There was not desaturation events in both group. CONCLUSIONS : Under careful monitoring the use of propofol for sedation during RFA is superior to midazolam/ pentazocine in patients with liver neoplasms. M1235 Deguelin As a New Therapeutic Candidate for the Treatment of Hepatocellular Carcinoma Ju-Hee Lee, Soon-Sun Hong, Seok Jeong, Don Haeng Lee, Hyung Gil Kim, Yong Woon Shin, Young Soo Kim Deguelin is a rotenoid of the flavonoid family isolated from several plant species including Mundulea sericea (Leguminosae) and a naturally occurring insecticide. It has previously been reported to have the potential as a chemopreventive agent against skin, mammary, colon, and lung cancer. The effects of deguelin on proliferation and apoptosis of hepatic cancer cells were assessed by the MTT assay and flow cytometric analysis. The growth of hepatic cancer cells (HepG2, Huh7, and SK-Hep1) was inhibited by deguelin in a dose-dependent manner. HepG2 cells of all the cell lines were the most sensitive to deguelin (IC 50 = 0.62 μM). The proportions of sub-G1 apoptotic cells increased 4.94 to 37.77 % by deguelin (0.01 - 10 μM) treatment for 3 days in HepG2 cells. The effects of deguelin on anti- angiogenesis of HepG2 cells were assessed using the western blot and RT-PCR analysis. Treatment of HepG2 cells with deguelin for 16 hours under hypoxia conditions reduced the expression of hypoxia-inducible factor 1α (HIF-1α) protein and vascular endothelial growth factor (VEGF) mRNA in a dose-dependent manner. To investigate whether deguelin shows antiangiogenic activities, we performed In Vitro and In Vivo angiogenesis assays. In tube formation assay, deguelin remarkably reduced the capillary network formation of HUVECs on Matrigel beds. Also, deguelin markedly decreased the migration of HUVECs compared to control. Moreover, deguelin remarkably reduced angiogenesis on the CAM of chick embryos. These results suggest that deguelin is potentially useful as a chemotherapeutic agent in hepatocellular carcinoma. A-367 AGA Abstracts M1236 Linear EUS Assessment and EUS-Fine Needle Aspiration of Hilar Lesions Manu K. Nayar, Derek Manas, Mark Egan, Viney Wadehra, Kofi Oppong Introduction Optimal management of proximal biliary strictures depends on adequate tissue diagnosis of suspected malignant lesions. Sensitivity of ERCP brush cytology alone for the diagnosis of all malignant strictures is between 33% and 58 %. EUS guided FNA of these lesions is challenging and to date there have been only two published series (1,2). We report our experience of EUS guided FNA of hilar lesions in a tertiary referral centre. Methods All patients who underwent EUS guided FNA of hilar lesions between April 2004 to July 2007 were identified. These patients had a provisional finding of a hilar lesion either on radiological imaging or ERCP. The final diagnosis was determined by surgical pathology, results of EUS FNA or follow up. Results 33 patients underwent 38 procedures for hilar lesions during the study period. There were 17 males and 16 females with a mean age of 66.71 years (range = 40 - 79). All but one patient (who had a hilar node) had a suspected hilar stricture. All patients underwent had radiological imaging (CT or MRI). Five patients had repeat procedures and FNA was unsuccessful in one patient. The average number of passes was 2.16 (range = 1 -3). The cytological specimen was adequate in 24 patients. The cytological diagnosis was adenocarcinoma (11) and benign (6). Six patients had definite diagnosis of adenocarcinoma on biliary brushings (3) and surgical pathology (3). The mean follow up period was 24.83 months (range = 4 - 48). The overall sensitivity, specificity, positive predictive value, negative predictive value and accuracy of EUS-FNA were 76%, 100%, 100%, 32% and 72% respectively. If only adequate aspirates were included in the analysis, the values were 82%, 100%, 100%, 32% and 69% respectively. Conclusion EUS guided FNA is a useful modality in the final diagnosis of hilar lesions. The results of our study are similar to previously reported studies (1,2). However the low negative predictive value limits its ability to exclude malignancy reliably in patients with suspected disease. References 1. DeWitt J, Misra L, Le Blanc J et al. EUS-guided FNA of proximal biliary strictures after negative ERCP brush cytology results. Gastrointest Endosc. 2006 Sep; 64(3): 325-33 2. Fritscher Ravens A, Boering DC, Knoefel WT et al. EUS-guided fine-needle aspiration of suspected hilar cholangiocarcinoma in potentially operable patients with negative brush cytology. Am J Gastroenterol. 2004 Jan;99(1):45 M1237 Dopamine D1 and D2 Receptor Subtypes Are Involved in Enteric Neural Control of Chloride and Bicarbonate Secretion in Guinea Pig Duodenum Yun Xia, Xiyu Wang, Guo-Du Wang, Fie Zou, Guijun Fei, Sumei Liu, Mei-Hua Qu, Xiaohong Sun, Wei Ren, Jackie D. Wood Dopamine is postulated to be a mediator in enteric neural regulation of intestinal secretion. We reported earlier that stimulation of the dopamine D1 receptor subtype hyperpolarized the membrane potential in S and AH type neurons and activation of the D2 receptor depolarized the membrane potential and enhanced excitability in both kinds of neurons in the guinea pig ileum (Neurogastroenterol Mot 2007, 19:41). The present study used immunofluorescence and Western blot analysis to study the distribution and chemical codes for neurons that expressed D1 and/or D2 immunoreactivity (IR) in the submucosal plexus. Immunoreactivity for both D1 and D2 receptors was expressed widely in enteric neurons, including ChAT-IR, calretinin-IR and calbindin-IR neurons in the submucosal and myenteric plexuses. Western Blots confirmed expression for both receptor subtypes. Muscle-stripped duodenal preparations were mounted in Ussing flux chambers for the recording of short- circuit current (Isc), which is an indicator of chloride secretion. The pH-stat method was used to study effects of dopaminergic stimulation on bicarbonate (HCO3 - ) secretion in Ussing chambers. Pharmacological agents were applied in the serosal compartment of the chamber. Mucosal HCO3 - secretion was suppressed in a concentration-dependent manner by the D1 receptor agonist, A68930, with an IC 50 of 5.67±3.24 µM (P<0.05, preps from 10 animals). Chloride secretion also was suppressed in concentration-dependent manner by A68930 with an IC 50 of 4.79±2.52µM (P<0.05, n=10 animals). The inhibitory action of A68930 was reversed by pre-treatment with the D1 receptor antagonist, SKF83566 (20- 40µM; P<0.05, n=6 animals). Application of the D2 receptor agonist, quinpirole (10-30 µM) stimulated both HCO3 - and Cl - secretion (P<0.05). The stimulatory action of quinpirole was enhanced by pretreatment with idazoxan, an adrenergic alpha-2 receptor antagonist (10µM) in all preps from 6 animals. Pretreatment with the D2 receptor antagonist, L741626 (20µM) suppressed quinpirole-evoked HCO3 - and Cl - secretion (P<0.05) in all preps from 6 animals. The presence of 0.5-µM tetrodotoxin in the serosal, but not mucosal compartment of the chamber, abolished the inhibitory action of A68930 and the stimulatory action of quinpirole on both HCO3 - and Cl - . The results are consistent with evidence that enteric secretomotor neurons express both dopaminergic D1 and D2 receptor subtypes in guinea pig duodenum. Stimulation of the D1 subtype suppresses neurogenic secretion.Stimulation of the D2 subtype enhances secretion for both HCO3- and Cl - . (Supported by NIH RO1 DK37238, RO1 DK-068258 and KO8 DK060468) M1238 Peptide YY Acts Through Y2 Receptors to Inhibit Stimulated Colonic Motor Function in Conscious Mice Lixin Wang, Guillaume Gourcerol, Pu-Qing Yuan, Mulugeta Million, Muriel H. Larauche, Yvette Tache Background: While peptide YY (PYY) is a well established inhibitor of intestinal secretion and gastric motility, little is known on PYY influence on colonic function In Vivo. Aim: To characterize PYY action on stimulated colonic function in conscious mice. Methods: Fecal pellet output (FPO) and diarrhea were monitored in adult male C57BL/6 mice in response to PYY, neuropeptide Y (NPY), Y1 agonists ([Leu31,Pro34]NPY, [Pro34]PYY), or Y2 agonist (PYY3-36) injected intraperitoneally (IP) before novelty or restraint, or IP injection of 5- hydroxytryptophan (5-HTP) or bethanechol; Y2 antagonist, BIIE-0246 was injected IP before PYY or PYY3-36 plus novelty stress or 5-HTP. Distal colonic contractions were recorded by a miniaturised-pressure transducer inserted 2 cm past the anus of mice injected IP with AGA Abstracts