- Email: [email protected]
Expression of thymidine phosphorylase and vascular endothelial growth factor in carcinoma of the papilla of vater
April 2000
examine the prognostic relevance with regard to survival. Results: Statistically significant hazard ratios (95%CI) were found for: the presence of goblet cell carcinoids in the appendix: 5.60 (1.07-29.37; p=0.04); nonmutated p53 expression: 3.29 (1.35-8.01; p=0.OOO9); MIBI: 3.15 (1.337.43; p=0.0009). The presence of liver metastases at diagnosis did not reach significance: 3.00 (1.0-9.05; p=0.051). Expression ofp53 and MIBI were closely related. The other variables failed to prove as independant prognostic factors influencing survival. Conclusion: Histologic proof of an increased proliferative activity and atypical appendiceal carcinoids seem to be associated with an unfavourable prognosis in patients with metastatic neuroendocrine GEP tumors.
2746 EXPRESSION OF THYMIDINE PHOSPHORYLASE AND VASCULAR ENDOTHELIAL GROWTH FACTOR IN CARCINOMA OF THE PAPILLA OF VATER. Bin Zhao, Kimura Wataru, Masatoshi Makuuchi, Shyoki Yazaki, Tokyo Univ Sch of Medicine, Tokyo, Japan; Yamagata Univ Sch of Medicine, Yamagata, Japan; T; Tohoku Univ Sch of Medicine, Tokyo, Japan. Background. Thymidine phosphorylase (TP) and vascular endothelial growth factor (VEGF) are important angiogenic factors which have been reported to be associated with tumor progression in malignant tumors. There has been no study of expression of angiogenic factors and neovascularization in carcinoma of the papilla of Vater. We investigated the expression of both TP and VEGF and the correlation with neovascularization and clinicopathological factors as well as clinical outcome in this malignancy. Methods. Fifty nine cases of carcinoma of the papilla of Vater were studied. All patients were surgically-treated at Department of Surgery, the University of Tokyo, from 1980 to 1997. There were 37 man and 22 women. The average age was 64 years (range 42 to 85). Tumor staging was according to TNM classification. An immunohistochemical method was used to evaluate the expression ofTP and VEGF as well as to quantify microvessel density (MVD) with formalin-fixed paraffin-embedded sections. MVD was defined as the microvessel count in a field with the most highest density of microvessels under a high magnification (about I ILmz in size). Results. TP expression was demonstrated in tumor cells in 62.7 % (37/59) of the cases. The expression was mainly cytoplasmic. A significant correlation was observed between TP expression and regional lymph node metastasis (p=0.006). TP-positive tumors were more advanced in stage as compared with TP-negative tumors (p=O.03). Moreover, the expression of TP correlated significantly with a poor prognosis (p=0.023). As regard to VEGF, 72.9% (43/59) of the cases was positive. The expression of VEGF was significantly correlated with venous invasion (p=0.042), and tumor ulceration (p=0.05). However, no survival significance was found (p=0.750). MVD in TP-positive tumors (27.6::':: 10.1) was significantly higher than that in TP-negative tumors (20.4::':: 10.0, p=O.OI). VEGF alone was not correlated with MVD. However, when TP and VEGF were combined, tumors positive for both had a significantly higher MVD (27.7::'::9.6) than that in tumors negative for both (18.2::'::8.5, p=O.OII). Conclusions. In carcinoma of the papilla of Vater, TP enhances tumor progression, and is a useful prognostic factor. Neovascularization in this malignancy is regulated by both TP and VEGF.
2747 OMEGA·3 FATTY ACIDS (03FA) ENHANCE CHEMOSENSITIV· ITY OF THE COLON CARCINOMA CELL LINE CACO·2 TO 5·FLUOROURACIL (5FU). Bora Akoglu, Angela Jordan, Wolfgang F. Caspary, Juergen Stein, J W Goethe Univ, Frankfurt, Germany; 2nd Dept Medicine, JW Goethe Univ, Frankfurt, Germany. Background/Aim: The use of 5-Fluorouracil (5FU), a chemotherapeutic agent for the treatment of colorectal carcinoma, is limited because of increasing antitumor drug resistance. We have recently shown (Gastroenterology 1999, G1880). that fish oil rich in omega-S fatty acids (03FA) inhibits growth of colorectal tumor cells. The objective of the present investigation was to determine the effect of an 03FA containing lipid emulsion on growth and chemosensitivity to 5FU in the colon carcinoma cell line, Caco-2. Methods: Caco-2 cells were exposed to two different lipid emulsions (Omegaven-lO%®on fish oil basis (FO) containing 2.1 g/Ioo mL eicosapentanoic acid (EPA); Lipovenos-l O'io PLR® on basis of soybean oil (SO), no omega-3-fatty acids ), alone or in combination with 5FU. Cell proliferation was determined by cell counting (crystal violet assay) and BrdU uptake after 24, 48 and 72 h. Cell cycle was assessed by flow cytometry. Results: Treatment of Caco-2 cells with FO induced a time- and dose-dependent decrease in cell counts and BrdU uptake. Cell counts:[% control after 24 h: -10::'::8(*); 48 h: -33::'::7(*); 72 h: -53::'::4(*)], BrdU uptake: [% control after 24 h: -67::'::2(**); 48 h: -86::':: 1(**); 72 h: -88::'::1(**)] at 15.5 ILI/loo ILl medium corresponding to 100 ILM EPA. Incubation with SO resulted in a less prominent effect. Cell counts [ % control after 24 h: -4::'::8(ns); 48 h: -1::'::12(*); 72 h: -23::'::6(*)], BrdV uptake: [% control after 24 h: -14::':: 11(*); 48 h: -64::'::8 (**); 72 h: -65±7(**)] at the same dose [(ns)=not significant, (*)=p<0.05, (**)=p
AGAA515
stance vs. combination). Conclusions: Fish oil based lipid emulsion (FO) has a potent antiproliferative effect on the colon carcinoma cell line Caco-2, due to cell cycle arrest. Addition of 5FU has an additive and/or synergistic growth inhibitory effect. (Supported by the Else Kroner Fresenius Stiftung)
2748 PHASE IIII TRIAL WITH lllIN·PENTETREOTIDE IN PATIENTS WITH ADVANCED MALIGNANCIES. Athanassios Argiris, Kathy Peccerillo, John R. Murren, Eugene Cornelius, Irvin M. Modlin, Yale Vniv Sch of Medicine, New Haven, CT. Radiolabeled somatostatin analogs can be utilized to deliver therapeutic doses of radiation to neuroendocrine and other tumors expressing somatostatin receptors in high density. We conducted a phase 1111 trial with 111 Indium-labeled DTPA-D-Phe-I-octreotide IIIn-pentetreotide) in patients (pts) with advanced malignancies with positive uptake on baseline diagnostic IIIIn-pentetreotide scan. 1llln-pentetreotide was administered intravenously every 3 weeks for a total of 4 cycles. at 3 dose levels: 160, 225, and 300 mCi/cycie. From 12196 to 8199 30 pts were enrolled: 25 with carcinoid or other gastroenteropancreatic endocrine tumors (GEPT), I small-cell lung cancer, I chordoma, I Hodgkin's disease, I thymoma. and I giant cell tumor (dose levels 1111111 : 6/4/20 pts). Mean age was 52 (range 27- 78); 18 malesl12 females. All pts were symptomatic with extensive metastatic disease; 11130 had prior chemotherapy. 4 pts who derived clinical benefit received 2 additional cycles (maximum cumulative dose 1.800 mCi). A total of 123 cycles of 111- In-pentetreotide have been delivered. Mild/moderate myelosuppression was common, and sometimes prolonged. At dose level III grade I or 2 thrombocytopenia developed in 11120 pts (55%), grade 3 leukopenia in 2 pts (10%), and grade I or 2 leukopenia in 6 pts (30%). Other non-hematologic toxicities, possibly unrelated to therapy, included acute renal failure due to dehydration (1), reversible neuropathy (1), psoriasis flare (1), and new onset diabetes (1). 6/23 evaluable pts with GEPT (26%) had symptomatic improvement (dose levels II and III), and 15/23 (65%) had stable symptomatology; 2 minor radiographic responses (dose levels II and III) and I partial response (PR) (level III) were observed. 5/18 evaluable pts (28%) had a > 50% decrease of at least one hormonal marker. In pts with other malignancies (N=5), I pt with Hodgkin's disease achieved a PR (level III). Radiographic responses were delayed, occurring 2-5 months after treatment initiation. Median time to progression was 6 months. IIIIn-pentetreotide could be escalated to a cumulative dose of at least 1,200 mCi with acceptable hematologic toxicities, and produced clinical benefit in patients with endstage malignancies.
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NFKB INHIBITION IN PANCREATIC CARCINOMA CELLS TRIGGERS APOPTOSIS INDUCED BY VP16 OR ADRIAMYCIN. Alexander Arlt, Anke Huenermann-Jansen, Ulrich R. Foelsch, Wolfgang E. Schmidt, Heiner Schaefer, Ist Dept of Medicine, Kiel, Germany; Dept of Medicine I. Bochum, Germany. Treatment of cancer with genotoxic agents is hampered by resistance to apoptosis exhibited by many tumors, including those of the gastrointestinal tract. Since NFKB mediated cellular responses are supposed to confer anti-apoptotic properties we investigated whether blockade of NFKB activity enhances the cytoxic effect of etoposide (VPI6) or adriamycin on pancreatic carcinoma cells. Amongst various pancreatic cancer cell lines, some (BxPC-3, PT45-1) were highly susceptible to apoptosis induced by VPI6 and adriamycin, whereas others were less sensitive or almost resistant (Capan-I, 818-4). These distinct apoptotic responses were verified by annexin-VIPI labelling and FACS analysis as well as by PARP cleavage analysis. Gel shift assays demonstrated that treatment with VPI6 (20 ILM), but not with adriamycin (0,3 ILM), elicits a rapid increase (I h) in NFKB binding activity in all cell lines, but the duration of this NFKBresponse and its basal activity significantly varied. In BxPc3- and PT45-1 cells, basal NFKB binding was nearly undetectable and the VPl6-induced activity rapidly declined. In Capan-l- and 818-4 cells, basal NFKB activity was strongly elevated and the period of NFKB activation lasted up to 24 h. By means of NFKB-Iuciferase assay, this distinct pattern of NFKB activity could be verified on the level of its transactivating capacity. Blockade of NFKB with agents that interfere with NFKB activation at distinct levels (Gliotoxin, MG132 and Sulfasalazine) not only abolished NFKB activation but also enhanced the apoptotic effects of VPI6 and adriamycin - particularly in those cell lines being resistant to these drugs. Our results indicate that the lack of a cytotoxic effect of chemotherapy in some tumors is due to constitutive NFKB activity rather than a transient induction of NFKB. Thus, blockade of NFKB activity by some well established immunosuppressive drugs obviously reduces resistance of GI-cancer cells.(Supported by the Deutsche Forschungsgemeinschaft: SFB 415,and by the IZKF: Immunobiology of Malignant Disease)
examine the prognostic relevance with regard to survival. Results: Statistically significant hazard ratios (95%CI) were found for: the presence of goblet cell carcinoids in the appendix: 5.60 (1.07-29.37; p=0.04); nonmutated p53 expression: 3.29 (1.35-8.01; p=0.OOO9); MIBI: 3.15 (1.337.43; p=0.0009). The presence of liver metastases at diagnosis did not reach significance: 3.00 (1.0-9.05; p=0.051). Expression ofp53 and MIBI were closely related. The other variables failed to prove as independant prognostic factors influencing survival. Conclusion: Histologic proof of an increased proliferative activity and atypical appendiceal carcinoids seem to be associated with an unfavourable prognosis in patients with metastatic neuroendocrine GEP tumors.
2746 EXPRESSION OF THYMIDINE PHOSPHORYLASE AND VASCULAR ENDOTHELIAL GROWTH FACTOR IN CARCINOMA OF THE PAPILLA OF VATER. Bin Zhao, Kimura Wataru, Masatoshi Makuuchi, Shyoki Yazaki, Tokyo Univ Sch of Medicine, Tokyo, Japan; Yamagata Univ Sch of Medicine, Yamagata, Japan; T; Tohoku Univ Sch of Medicine, Tokyo, Japan. Background. Thymidine phosphorylase (TP) and vascular endothelial growth factor (VEGF) are important angiogenic factors which have been reported to be associated with tumor progression in malignant tumors. There has been no study of expression of angiogenic factors and neovascularization in carcinoma of the papilla of Vater. We investigated the expression of both TP and VEGF and the correlation with neovascularization and clinicopathological factors as well as clinical outcome in this malignancy. Methods. Fifty nine cases of carcinoma of the papilla of Vater were studied. All patients were surgically-treated at Department of Surgery, the University of Tokyo, from 1980 to 1997. There were 37 man and 22 women. The average age was 64 years (range 42 to 85). Tumor staging was according to TNM classification. An immunohistochemical method was used to evaluate the expression ofTP and VEGF as well as to quantify microvessel density (MVD) with formalin-fixed paraffin-embedded sections. MVD was defined as the microvessel count in a field with the most highest density of microvessels under a high magnification (about I ILmz in size). Results. TP expression was demonstrated in tumor cells in 62.7 % (37/59) of the cases. The expression was mainly cytoplasmic. A significant correlation was observed between TP expression and regional lymph node metastasis (p=0.006). TP-positive tumors were more advanced in stage as compared with TP-negative tumors (p=O.03). Moreover, the expression of TP correlated significantly with a poor prognosis (p=0.023). As regard to VEGF, 72.9% (43/59) of the cases was positive. The expression of VEGF was significantly correlated with venous invasion (p=0.042), and tumor ulceration (p=0.05). However, no survival significance was found (p=0.750). MVD in TP-positive tumors (27.6::':: 10.1) was significantly higher than that in TP-negative tumors (20.4::':: 10.0, p=O.OI). VEGF alone was not correlated with MVD. However, when TP and VEGF were combined, tumors positive for both had a significantly higher MVD (27.7::'::9.6) than that in tumors negative for both (18.2::'::8.5, p=O.OII). Conclusions. In carcinoma of the papilla of Vater, TP enhances tumor progression, and is a useful prognostic factor. Neovascularization in this malignancy is regulated by both TP and VEGF.
2747 OMEGA·3 FATTY ACIDS (03FA) ENHANCE CHEMOSENSITIV· ITY OF THE COLON CARCINOMA CELL LINE CACO·2 TO 5·FLUOROURACIL (5FU). Bora Akoglu, Angela Jordan, Wolfgang F. Caspary, Juergen Stein, J W Goethe Univ, Frankfurt, Germany; 2nd Dept Medicine, JW Goethe Univ, Frankfurt, Germany. Background/Aim: The use of 5-Fluorouracil (5FU), a chemotherapeutic agent for the treatment of colorectal carcinoma, is limited because of increasing antitumor drug resistance. We have recently shown (Gastroenterology 1999, G1880). that fish oil rich in omega-S fatty acids (03FA) inhibits growth of colorectal tumor cells. The objective of the present investigation was to determine the effect of an 03FA containing lipid emulsion on growth and chemosensitivity to 5FU in the colon carcinoma cell line, Caco-2. Methods: Caco-2 cells were exposed to two different lipid emulsions (Omegaven-lO%®on fish oil basis (FO) containing 2.1 g/Ioo mL eicosapentanoic acid (EPA); Lipovenos-l O'io PLR® on basis of soybean oil (SO), no omega-3-fatty acids ), alone or in combination with 5FU. Cell proliferation was determined by cell counting (crystal violet assay) and BrdU uptake after 24, 48 and 72 h. Cell cycle was assessed by flow cytometry. Results: Treatment of Caco-2 cells with FO induced a time- and dose-dependent decrease in cell counts and BrdU uptake. Cell counts:[% control after 24 h: -10::'::8(*); 48 h: -33::'::7(*); 72 h: -53::'::4(*)], BrdU uptake: [% control after 24 h: -67::'::2(**); 48 h: -86::':: 1(**); 72 h: -88::'::1(**)] at 15.5 ILI/loo ILl medium corresponding to 100 ILM EPA. Incubation with SO resulted in a less prominent effect. Cell counts [ % control after 24 h: -4::'::8(ns); 48 h: -1::'::12(*); 72 h: -23::'::6(*)], BrdV uptake: [% control after 24 h: -14::':: 11(*); 48 h: -64::'::8 (**); 72 h: -65±7(**)] at the same dose [(ns)=not significant, (*)=p<0.05, (**)=p
AGAA515
stance vs. combination). Conclusions: Fish oil based lipid emulsion (FO) has a potent antiproliferative effect on the colon carcinoma cell line Caco-2, due to cell cycle arrest. Addition of 5FU has an additive and/or synergistic growth inhibitory effect. (Supported by the Else Kroner Fresenius Stiftung)
2748 PHASE IIII TRIAL WITH lllIN·PENTETREOTIDE IN PATIENTS WITH ADVANCED MALIGNANCIES. Athanassios Argiris, Kathy Peccerillo, John R. Murren, Eugene Cornelius, Irvin M. Modlin, Yale Vniv Sch of Medicine, New Haven, CT. Radiolabeled somatostatin analogs can be utilized to deliver therapeutic doses of radiation to neuroendocrine and other tumors expressing somatostatin receptors in high density. We conducted a phase 1111 trial with 111 Indium-labeled DTPA-D-Phe-I-octreotide IIIn-pentetreotide) in patients (pts) with advanced malignancies with positive uptake on baseline diagnostic IIIIn-pentetreotide scan. 1llln-pentetreotide was administered intravenously every 3 weeks for a total of 4 cycles. at 3 dose levels: 160, 225, and 300 mCi/cycie. From 12196 to 8199 30 pts were enrolled: 25 with carcinoid or other gastroenteropancreatic endocrine tumors (GEPT), I small-cell lung cancer, I chordoma, I Hodgkin's disease, I thymoma. and I giant cell tumor (dose levels 1111111 : 6/4/20 pts). Mean age was 52 (range 27- 78); 18 malesl12 females. All pts were symptomatic with extensive metastatic disease; 11130 had prior chemotherapy. 4 pts who derived clinical benefit received 2 additional cycles (maximum cumulative dose 1.800 mCi). A total of 123 cycles of 111- In-pentetreotide have been delivered. Mild/moderate myelosuppression was common, and sometimes prolonged. At dose level III grade I or 2 thrombocytopenia developed in 11120 pts (55%), grade 3 leukopenia in 2 pts (10%), and grade I or 2 leukopenia in 6 pts (30%). Other non-hematologic toxicities, possibly unrelated to therapy, included acute renal failure due to dehydration (1), reversible neuropathy (1), psoriasis flare (1), and new onset diabetes (1). 6/23 evaluable pts with GEPT (26%) had symptomatic improvement (dose levels II and III), and 15/23 (65%) had stable symptomatology; 2 minor radiographic responses (dose levels II and III) and I partial response (PR) (level III) were observed. 5/18 evaluable pts (28%) had a > 50% decrease of at least one hormonal marker. In pts with other malignancies (N=5), I pt with Hodgkin's disease achieved a PR (level III). Radiographic responses were delayed, occurring 2-5 months after treatment initiation. Median time to progression was 6 months. IIIIn-pentetreotide could be escalated to a cumulative dose of at least 1,200 mCi with acceptable hematologic toxicities, and produced clinical benefit in patients with endstage malignancies.
e
2749
NFKB INHIBITION IN PANCREATIC CARCINOMA CELLS TRIGGERS APOPTOSIS INDUCED BY VP16 OR ADRIAMYCIN. Alexander Arlt, Anke Huenermann-Jansen, Ulrich R. Foelsch, Wolfgang E. Schmidt, Heiner Schaefer, Ist Dept of Medicine, Kiel, Germany; Dept of Medicine I. Bochum, Germany. Treatment of cancer with genotoxic agents is hampered by resistance to apoptosis exhibited by many tumors, including those of the gastrointestinal tract. Since NFKB mediated cellular responses are supposed to confer anti-apoptotic properties we investigated whether blockade of NFKB activity enhances the cytoxic effect of etoposide (VPI6) or adriamycin on pancreatic carcinoma cells. Amongst various pancreatic cancer cell lines, some (BxPC-3, PT45-1) were highly susceptible to apoptosis induced by VPI6 and adriamycin, whereas others were less sensitive or almost resistant (Capan-I, 818-4). These distinct apoptotic responses were verified by annexin-VIPI labelling and FACS analysis as well as by PARP cleavage analysis. Gel shift assays demonstrated that treatment with VPI6 (20 ILM), but not with adriamycin (0,3 ILM), elicits a rapid increase (I h) in NFKB binding activity in all cell lines, but the duration of this NFKBresponse and its basal activity significantly varied. In BxPc3- and PT45-1 cells, basal NFKB binding was nearly undetectable and the VPl6-induced activity rapidly declined. In Capan-l- and 818-4 cells, basal NFKB activity was strongly elevated and the period of NFKB activation lasted up to 24 h. By means of NFKB-Iuciferase assay, this distinct pattern of NFKB activity could be verified on the level of its transactivating capacity. Blockade of NFKB with agents that interfere with NFKB activation at distinct levels (Gliotoxin, MG132 and Sulfasalazine) not only abolished NFKB activation but also enhanced the apoptotic effects of VPI6 and adriamycin - particularly in those cell lines being resistant to these drugs. Our results indicate that the lack of a cytotoxic effect of chemotherapy in some tumors is due to constitutive NFKB activity rather than a transient induction of NFKB. Thus, blockade of NFKB activity by some well established immunosuppressive drugs obviously reduces resistance of GI-cancer cells.(Supported by the Deutsche Forschungsgemeinschaft: SFB 415,and by the IZKF: Immunobiology of Malignant Disease)