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199Effects of a new oncolytic adenovirus (XVIR) in human prostate cancer cell lines and in xenograph mouse model
197 PROGNOSTIC SIGNIFICANCE OF I N F E R I O R VENA CAVA INVASION IN 764 T3B A N D T 3 C R E N A L T U M O U R S
INFRARED & MASS S P E C T R A L I M A G I N G OF PROSTATE C A N C E R CELLS
Bensalah K. ~, Guille F. ~, De la Taille A. 2, Ficarra V.3, Cincolo L. 4, Chopin D. 2, Abbou C.C. z, Altieri V.4, Artibani W. 3, Tostain j.5, Lobel B. 6, Patard j.j.6
Brown M. 1, Gazi E. 1, Gardner p.2, Dwyer j.2, Lockyer N. 2, Vickerman j.2 Miyan j.3, Hart C. ~, Clarke N. 1
1H6pital Ponchaillou, Dept. of Urology, Rennes, France, 2Henri Mondor Hospital, Dept. of Urology, Creteil, France, 3Verona University Hospital, Dept. of Urology, Verona, Italy, 4Naples University Hospital, Dept. of Urology, Naples, Italy, 5Saint Etienne University Hospital, Dept. of Urology, Saint Etienne, France, 6Rennes University Hospital, Dept. of Urology, Rennes, France INTRODUCTION & O B J E C T I V E S : To evaluate the prognostic significance of IVC invasion in renal tumours invading the venous system (T3b and c). MATERIAL & M E T H O D S : 764 patients with renal vein invasion, including 155 (3.9%) with IVC invasion were selected from a 4040 patients multicentre database. T3b tumours with or without IVC invasion were compared. Additionally, the following criteria were evaluated for their prognostic value: age, sex, IVC invasion, perinephric fat, adrenal, nodal or distant metastatic invasion. Uni and multivariate analysis were performed by using Kaplan Meier and Cox methods respectively. RESULTS: Tumours with IVC invasion were greater in size, were more likely to be symptomatic (p<0.001) and to invade adrenal gland (p :0.02). Compared to other pT3b renal tumours, patients with IVC invasion had a poorer performance status (p :0.01). Age, sex, histologic subtype, fat, nodal or distant metastatic invasion rates did not differ between the 2 groups. In univariate analysis: tumour size, ECOG Performance Status, symptoms, fat, adrenal, nodal, distant metastatic and IVC invasion (p<0.001) as well as histologic subtype (p:0.02) were significant prognostic factors. In multivariate analysis, nodal and metastatic invasion (p<0.001), fat invasion (p:0.001); symptoms at diagnosis (p:0.01), and IVC invasion (p:0.04) remained independent prognostic factors. CONCLUSIONS: Renal tumours with IVC invasion differ from other T3b tumours by local invasiveness, Symptoms at diagnosis and altered performance status. Contrary to the general belief, patients with IVC invasion seem to have a poorer outcome compared to patients with a less extensive venous invasion.
~Cancer Research UK Paterson Institute, Prompt Genitu-Urinary Cancer Research Group, Manchester, United Kingdom, 2University of Manchester, School of Chemical Engineering and Analytical Science, Manchester, United Kingdom, 3University of Manchester, Biomolecular Sciences, Manchester, United Kingdom INTRODUCTION & OBJECTIVES: IR microspectroscopy measures the vibrational
modes of functional groups of biomolecules as IR light is transmitted through micron-scale regions of the cell. IR spectral imaging can be applied to the study of single cells to reveal the spatial distribution and concentration profiles of lipid, protein and phosphate domains. Optimisatiun to both spatial and spectral resolution is realised using synchrotron (SR) based IR radiation, which is 1000 times more brilliant than IR laboratory sources. Here we apply SRIR imaging technology to the study of dynamic cellular events, cytokinesis and cell motility in the PC-3 prostate cancer cell lines. Additionally, we have utilised imaging time-of-flight secondary ion mass spectrometry (ToF-SIMS) to acquire biochemical images of the PC-3 cell. This technique allows the logalisation and identification of target molecular and ionic species from the outermost layers of a sub-micrometer scale surface allowing chemical analysis of different cellular sections following freeze-fracture. MATERIAL & METHODS: PC-3 cells cultured in Ham's F12 media upon MirrlR slides and fixed in 4% formalin were analysed by SR-IR. For the ToF-SIMS study, PC-3 cells were cultured onto silicon and flash frozen as a sandwich for freeze-fracture. The specimen was freeze-dried and analysed under vacuum in the ToF-SIMS instrument RESULTS: SR-IR chemical images of the cytokinetic cell showed evidence for additional membrane at the cleavage furrow (depicted by the methylene and methyl modes), cytoplasmic t~anslocation of membrane rich organelles (depicted by the lipid carbonyl ester vibrational mode) and contractile ring assembly at the midbody (depicted by the amide I and II modes). SR-IR images of the lipid intensity profiles in motile cells, suggested differences in the plasma membrane composition, compared to stationary ceils, which may be associated with changes in the plasma membrane fluidity during cell locomotion. ToF-SIMS analysis of the fractured PC-3 demonstrates localisation of various species including K, Cu, Ca and Mg within the cytoplasm. Also, mass signals corresponding to different phospholipids were used to elucidate different fracture planes in the cell image. This allowed for confirmation of Cn accumulation (required for activation of the angiogenic switch) within the cell cytoplasm and of lower concentrations in the basolateral leaf of the plasma membrane. CONCLUSIONS: SR-IR imaging can depict localisation of structural components as well as cellular processes within cells from prostate and other cancers. ToF- SIMS imaging of the fractured cell provides unique insight into intercellular elemental ion localisations, which are present at millimolar concentrations and are vital to cell physiology.
199
200
EFFECTS OF A N E W ONCOLYTIC ADENOVIRUS (XVIR) IN H U M A N PROSTATE C A N C E R C E L L LINES AND IN X E N O G R A P H MOUSE MODEL
E F F E C T OF R O F E C O X I B AND C E L E C O X I B USAGE ON PROSTATE C A N C E R R I S K - A POPULATION BASED CASE- C O N T R O L STUDY
Treiber U. 1, Mantwill K. 2, K6hler Vargas N~2, Berushansen A. 2, Paul R. 1, Van Randenborgh H. 1, Kiibler H. 1, Zantl N. ~, Hartung R. 1, G/insbacher B. 2, Holm ES. 2 1Technische Universitiit Miinchen, Urology, Munich, Germany, 2Technische Universitiit Miinchen, Experimental Oncology, Munich, Germany & OBJECTIVES: Treatment of locally advanced and especially metastatic hormone refractory prostate cancer is a major therapeutic challenge due to development of multidrug resistance phenotype. Improvement of current therapy modalities by addition of oncolytic virotherapy is an innovative approach for this type of cancer. INTRODUCTION
MATERIAL & METHODS: The cytolytic effects of a new developed E 1-minus, YB-1 dependent, replicating oncolytic adenovirus ("Xvir"; developed by XVir Therapeutics GmbH, Munich, Germany: M.K., H.RS.) were assessed in three human prostate cancer cell lines (LNCaP, DU145, PC3) by monolayer assay in 6-well plates (1-100 plaque-forming units; pfu/cell). Furthermore, virus and low concentrations (IC20: 20% growth inhibition, 80% cell survival) of cytostatic agents (irinotecan, trichostatin A) were applicated simultaneously. Additionally, a xenograph mouse model with DU145 tumours was established and effects of intratumoral application of Xvir were investigated in vivo. RESULTS: Virus application resulted in complete cytolysis in all three cell lines. Combined application of virus and chemotherapeutic agents lead to synergistic cytolytic effects. These effects were mediated by nuclear translocation of transcription factor YB-I, which facilitates virus replication (Holm 2002, JBC 277: 10427-10434; Holm 2004, Cancer Res 64: 322-328). Furthermore, strong tumour growth inhibition and tumour regression was observed in vivo. CONCLUSIONS: Implementation of potent oncolytic adenoviruses in current concepts for prostate cancer treatment represents the beginning of a new era of therapeutic tools in this tumour entity. The recently designed, replicative adenovims Xvir (developed by XVir Therapeutics GmbH, Munich, Germany: M.K., H.P.S.) showed highly effective cytolytic capacities, especially in combination with cytostatic agents and established the crucial role of transcription factor YB- 1 in adenoviral replication. Therefore, combination of adenoviral therapy with cytostatic agents or radiation delineates a novel therapeutic strategy for prostate cancer, which should be further investigated in vivo and in clinical trials.
European Urology Supplements 4 (2005) No. 3, pp. 52
Murtola %1, Tammela T.2, Mfifittfinen L ) , Auvinen A. 1, Screening for prostate cancer with PSA-testing, Helsinki, Finland 1University of Tampere, School of Public Health, Epidemiology, Tampere, Finland, 2Tampere University Hospital, Urology, Tampere, Finland, 3The Finnish Cancer Registry, Cancer Statistics, Helsinki, Finland I N T R O D U C T I O N & OBJECTIVES: Selective cyclo-oxygenase (COX)-2inhibiting drugs rofecoxib and celecoxib have shown promise in treating and in some cases preventing prostate cancer. In vitro and some in vivo animal studies suggest these drugs to increase apoptosis and decrease angiogenesis in prostate cancer cells and tissues. The objective of this study was to evaluate the effect of coxib usage on prostate cancer risk at the population level. M A T E R I A L & M E T H O D S : A population-based case-control study was conducted using the Finnish national registries. Cases, all men diagnosed with prostate cancer in Finland in 2002, were identified from the Finnish Cancer Registry. Controls, matched for age and residence area at the selection, were obtained from the Finnish Population Register Centre. Information on rofecoxib and celecoxib usage was obtained from the comprehensive nationwide prescription database of the Social Insurance Institution of Finland. Total mg amounts of both drugs used during 2000-2002 until prostate cancer diagnosis were calculated for each person. Cross-tabulation and conditional logistic regression methods were used to calculate frequencies, ORs and 95% CIs for coxib usage. R E S U L T S : 3 879 cases and 3 865 controls were included. Of the cases 2.0 % used rofecoxib, 1.6 % used celecoxib and 0.2 % used both. For the controls the percentages were 2.1%, 1.7 % and 0.1%, respectively. Having used either coxib was not associated with a statistically significantly increased risk for prostate cancer; OR 0.916 for rofecoxib (95 % CI 0.723-1.160) and OR 0.938 for celecoxib (95 % CI 0.722-1.218). OR for having used both was 1.514 (95 % CI 0.6173.716). Effect of coxib usage on prostate cancer risk remained statistically nonsignificant in all subgroups categorized by the regularity of drug use.
CONCLUSIONS: Rofecoxib and celecoxib usage did not affect the risk of prostate cancer at the population level. The follow-up time, however, is short due to recent arrival of coxibs in Finland. The results require further confirmation, particularly from possible chemoprevention trials.
INFRARED & MASS S P E C T R A L I M A G I N G OF PROSTATE C A N C E R CELLS
Bensalah K. ~, Guille F. ~, De la Taille A. 2, Ficarra V.3, Cincolo L. 4, Chopin D. 2, Abbou C.C. z, Altieri V.4, Artibani W. 3, Tostain j.5, Lobel B. 6, Patard j.j.6
Brown M. 1, Gazi E. 1, Gardner p.2, Dwyer j.2, Lockyer N. 2, Vickerman j.2 Miyan j.3, Hart C. ~, Clarke N. 1
1H6pital Ponchaillou, Dept. of Urology, Rennes, France, 2Henri Mondor Hospital, Dept. of Urology, Creteil, France, 3Verona University Hospital, Dept. of Urology, Verona, Italy, 4Naples University Hospital, Dept. of Urology, Naples, Italy, 5Saint Etienne University Hospital, Dept. of Urology, Saint Etienne, France, 6Rennes University Hospital, Dept. of Urology, Rennes, France INTRODUCTION & O B J E C T I V E S : To evaluate the prognostic significance of IVC invasion in renal tumours invading the venous system (T3b and c). MATERIAL & M E T H O D S : 764 patients with renal vein invasion, including 155 (3.9%) with IVC invasion were selected from a 4040 patients multicentre database. T3b tumours with or without IVC invasion were compared. Additionally, the following criteria were evaluated for their prognostic value: age, sex, IVC invasion, perinephric fat, adrenal, nodal or distant metastatic invasion. Uni and multivariate analysis were performed by using Kaplan Meier and Cox methods respectively. RESULTS: Tumours with IVC invasion were greater in size, were more likely to be symptomatic (p<0.001) and to invade adrenal gland (p :0.02). Compared to other pT3b renal tumours, patients with IVC invasion had a poorer performance status (p :0.01). Age, sex, histologic subtype, fat, nodal or distant metastatic invasion rates did not differ between the 2 groups. In univariate analysis: tumour size, ECOG Performance Status, symptoms, fat, adrenal, nodal, distant metastatic and IVC invasion (p<0.001) as well as histologic subtype (p:0.02) were significant prognostic factors. In multivariate analysis, nodal and metastatic invasion (p<0.001), fat invasion (p:0.001); symptoms at diagnosis (p:0.01), and IVC invasion (p:0.04) remained independent prognostic factors. CONCLUSIONS: Renal tumours with IVC invasion differ from other T3b tumours by local invasiveness, Symptoms at diagnosis and altered performance status. Contrary to the general belief, patients with IVC invasion seem to have a poorer outcome compared to patients with a less extensive venous invasion.
~Cancer Research UK Paterson Institute, Prompt Genitu-Urinary Cancer Research Group, Manchester, United Kingdom, 2University of Manchester, School of Chemical Engineering and Analytical Science, Manchester, United Kingdom, 3University of Manchester, Biomolecular Sciences, Manchester, United Kingdom INTRODUCTION & OBJECTIVES: IR microspectroscopy measures the vibrational
modes of functional groups of biomolecules as IR light is transmitted through micron-scale regions of the cell. IR spectral imaging can be applied to the study of single cells to reveal the spatial distribution and concentration profiles of lipid, protein and phosphate domains. Optimisatiun to both spatial and spectral resolution is realised using synchrotron (SR) based IR radiation, which is 1000 times more brilliant than IR laboratory sources. Here we apply SRIR imaging technology to the study of dynamic cellular events, cytokinesis and cell motility in the PC-3 prostate cancer cell lines. Additionally, we have utilised imaging time-of-flight secondary ion mass spectrometry (ToF-SIMS) to acquire biochemical images of the PC-3 cell. This technique allows the logalisation and identification of target molecular and ionic species from the outermost layers of a sub-micrometer scale surface allowing chemical analysis of different cellular sections following freeze-fracture. MATERIAL & METHODS: PC-3 cells cultured in Ham's F12 media upon MirrlR slides and fixed in 4% formalin were analysed by SR-IR. For the ToF-SIMS study, PC-3 cells were cultured onto silicon and flash frozen as a sandwich for freeze-fracture. The specimen was freeze-dried and analysed under vacuum in the ToF-SIMS instrument RESULTS: SR-IR chemical images of the cytokinetic cell showed evidence for additional membrane at the cleavage furrow (depicted by the methylene and methyl modes), cytoplasmic t~anslocation of membrane rich organelles (depicted by the lipid carbonyl ester vibrational mode) and contractile ring assembly at the midbody (depicted by the amide I and II modes). SR-IR images of the lipid intensity profiles in motile cells, suggested differences in the plasma membrane composition, compared to stationary ceils, which may be associated with changes in the plasma membrane fluidity during cell locomotion. ToF-SIMS analysis of the fractured PC-3 demonstrates localisation of various species including K, Cu, Ca and Mg within the cytoplasm. Also, mass signals corresponding to different phospholipids were used to elucidate different fracture planes in the cell image. This allowed for confirmation of Cn accumulation (required for activation of the angiogenic switch) within the cell cytoplasm and of lower concentrations in the basolateral leaf of the plasma membrane. CONCLUSIONS: SR-IR imaging can depict localisation of structural components as well as cellular processes within cells from prostate and other cancers. ToF- SIMS imaging of the fractured cell provides unique insight into intercellular elemental ion localisations, which are present at millimolar concentrations and are vital to cell physiology.
199
200
EFFECTS OF A N E W ONCOLYTIC ADENOVIRUS (XVIR) IN H U M A N PROSTATE C A N C E R C E L L LINES AND IN X E N O G R A P H MOUSE MODEL
E F F E C T OF R O F E C O X I B AND C E L E C O X I B USAGE ON PROSTATE C A N C E R R I S K - A POPULATION BASED CASE- C O N T R O L STUDY
Treiber U. 1, Mantwill K. 2, K6hler Vargas N~2, Berushansen A. 2, Paul R. 1, Van Randenborgh H. 1, Kiibler H. 1, Zantl N. ~, Hartung R. 1, G/insbacher B. 2, Holm ES. 2 1Technische Universitiit Miinchen, Urology, Munich, Germany, 2Technische Universitiit Miinchen, Experimental Oncology, Munich, Germany & OBJECTIVES: Treatment of locally advanced and especially metastatic hormone refractory prostate cancer is a major therapeutic challenge due to development of multidrug resistance phenotype. Improvement of current therapy modalities by addition of oncolytic virotherapy is an innovative approach for this type of cancer. INTRODUCTION
MATERIAL & METHODS: The cytolytic effects of a new developed E 1-minus, YB-1 dependent, replicating oncolytic adenovirus ("Xvir"; developed by XVir Therapeutics GmbH, Munich, Germany: M.K., H.RS.) were assessed in three human prostate cancer cell lines (LNCaP, DU145, PC3) by monolayer assay in 6-well plates (1-100 plaque-forming units; pfu/cell). Furthermore, virus and low concentrations (IC20: 20% growth inhibition, 80% cell survival) of cytostatic agents (irinotecan, trichostatin A) were applicated simultaneously. Additionally, a xenograph mouse model with DU145 tumours was established and effects of intratumoral application of Xvir were investigated in vivo. RESULTS: Virus application resulted in complete cytolysis in all three cell lines. Combined application of virus and chemotherapeutic agents lead to synergistic cytolytic effects. These effects were mediated by nuclear translocation of transcription factor YB-I, which facilitates virus replication (Holm 2002, JBC 277: 10427-10434; Holm 2004, Cancer Res 64: 322-328). Furthermore, strong tumour growth inhibition and tumour regression was observed in vivo. CONCLUSIONS: Implementation of potent oncolytic adenoviruses in current concepts for prostate cancer treatment represents the beginning of a new era of therapeutic tools in this tumour entity. The recently designed, replicative adenovims Xvir (developed by XVir Therapeutics GmbH, Munich, Germany: M.K., H.P.S.) showed highly effective cytolytic capacities, especially in combination with cytostatic agents and established the crucial role of transcription factor YB- 1 in adenoviral replication. Therefore, combination of adenoviral therapy with cytostatic agents or radiation delineates a novel therapeutic strategy for prostate cancer, which should be further investigated in vivo and in clinical trials.
European Urology Supplements 4 (2005) No. 3, pp. 52
Murtola %1, Tammela T.2, Mfifittfinen L ) , Auvinen A. 1, Screening for prostate cancer with PSA-testing, Helsinki, Finland 1University of Tampere, School of Public Health, Epidemiology, Tampere, Finland, 2Tampere University Hospital, Urology, Tampere, Finland, 3The Finnish Cancer Registry, Cancer Statistics, Helsinki, Finland I N T R O D U C T I O N & OBJECTIVES: Selective cyclo-oxygenase (COX)-2inhibiting drugs rofecoxib and celecoxib have shown promise in treating and in some cases preventing prostate cancer. In vitro and some in vivo animal studies suggest these drugs to increase apoptosis and decrease angiogenesis in prostate cancer cells and tissues. The objective of this study was to evaluate the effect of coxib usage on prostate cancer risk at the population level. M A T E R I A L & M E T H O D S : A population-based case-control study was conducted using the Finnish national registries. Cases, all men diagnosed with prostate cancer in Finland in 2002, were identified from the Finnish Cancer Registry. Controls, matched for age and residence area at the selection, were obtained from the Finnish Population Register Centre. Information on rofecoxib and celecoxib usage was obtained from the comprehensive nationwide prescription database of the Social Insurance Institution of Finland. Total mg amounts of both drugs used during 2000-2002 until prostate cancer diagnosis were calculated for each person. Cross-tabulation and conditional logistic regression methods were used to calculate frequencies, ORs and 95% CIs for coxib usage. R E S U L T S : 3 879 cases and 3 865 controls were included. Of the cases 2.0 % used rofecoxib, 1.6 % used celecoxib and 0.2 % used both. For the controls the percentages were 2.1%, 1.7 % and 0.1%, respectively. Having used either coxib was not associated with a statistically significantly increased risk for prostate cancer; OR 0.916 for rofecoxib (95 % CI 0.723-1.160) and OR 0.938 for celecoxib (95 % CI 0.722-1.218). OR for having used both was 1.514 (95 % CI 0.6173.716). Effect of coxib usage on prostate cancer risk remained statistically nonsignificant in all subgroups categorized by the regularity of drug use.
CONCLUSIONS: Rofecoxib and celecoxib usage did not affect the risk of prostate cancer at the population level. The follow-up time, however, is short due to recent arrival of coxibs in Finland. The results require further confirmation, particularly from possible chemoprevention trials.