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708 Dysregulation of cellular retinoic acid-binding protein (CRABP)-1 in renal cell carcinoma
708 DYSREGULATION PROTEIN (CRABP)-1 Schrader Hofmann
A.J.l, R. ’
‘University Braunschweig,
OF CELLULAR IN RENAL CELL
Hansen
W.*, Pfoertner
RETINOIC CARCINOMA
S.2, G&den
ACID-BINDING
U.2, Buer J.2, Heidenreich
709 VALUE OF PROGNOSTIC CARCINOMA
CYCLOOXYGENASE-2 INDICATORS
IN
Department of Urology, Marburg, Germany, of Cell Biology, Braunschweig, Germany
EXPRESSION RENAL
‘GBF
INTRODUCTION & OBJECTIVES: Retinoic acid (RA) is known to effect cell differentiation and proliferation and has been shown to inhibit or reverse the process of malignant transformation in several cell types. Although in vitro studies have been promising, clinical use of RA-derivates in renal cell carcinoma (RCC) did not meet first expectations. However, until today the mechanism of RA action and especially the role of the cellular retinoic acid-binding proteins (CRABP) are poorly understood. In this study. we quantified CRABP-1 mRNA expression in RCC and corresponding nonmalignant kidney tissue. Moreover, we overexpressed CRABP-1 in A498 RCC cells to disclose its potential influence on RA action in RCC. MATERIAL & METHODS: We used real-time RT-PCR methodology to investigate CRABP-1 expression in 14 RCC samples, 5 RCC cell lines (A498; CAKI-1, CAKI-2: HA-7, and LB1047) and the embryonal kidney cell line 293. Moreover, we cloned CRABP-1 to overexpress it in A498 cells via a CMV promoter. Afterwards, CRABP-1 expressing and n&e A498 cells were stimulated with at-RA and 9cis-RA; the antiproliferative effect of the RA-isotyps was evaluated in both cell types using 3H-thymidine-proliferation assays. RESULTS: The median tumour / kidney ratio for CRABP-1 expression was 0.0006 (range, 0.003.0.00002), displaying a significant downregulation of CRABP-1 in RCC. In fact, in tumour cell lines as well as in all 14 tumour samples, CRABP-1 was barely or not detectable. In contrast, CRABP-1 mRNA was expressed in embryonal 293 cells as well as in normal kidney tissue. Using proliferation assays to investigate the potential influence of CRABP-1 on the effect of RA stimulation in A498 cells, CRABP-1 expressing A498 displayed significant resistance to RA stimulation compared to nai+e A498 cells (40 and 72% at 1pM 9cis- and at-RA, respectively). CONCLUSIONS: Surprisingly, we were able to demonstrate a significant downregulation of CRABP-1 mRNA expression in all RCC cell lines as well as primary tumour samples compared to the corresponding normal kidney tissue. On the other hand, CRABP-1 overexpression in A498 RCC cells significantly inhibited RA associated antiproliferative actions. Thus, the dysregulation of CRABP-1 in RCC is probably not responsible for the limited efficiency of RA in RCC treatment.
‘Ajou University zAjou University
Choi M.K.‘,
KimYS.’
School of Medicine, Department of Urology, School of Medicine, Department of Pathology,
Suwon, South Korea, Suwon, South Korea
INTRODUCTION & OBJECTIVES: Cyclooxgenase (COX)-2 plays an important role in tumour cell proliferation, resistance to apoptosis, angiogenesis, and invasion in various cancers. However, the relationship between COX-2 expression and renal cell carcinoma (RCC) is not clear. The relationship between p53 expression and RCC is also still controversial. In this study, we investigated the expressions of COX-2 and p53 and related those to the clinicopathological variables in patients with conventional RCC. MATERIAL & METHODS: Formalin-fixed, paraffin-embedded tissue sections of conventional RCC from 92 patients who had undergone radical nephrectomy were stained immunohistochemically with specific antibodies against COX-2 and ~53. COX2 immunostaining was semi-quantitatively estimated based on proportion (percentage of positive cells) and intensity. COX-2 score was generated by multiplication of the values for the two parameters. P53 immunostaining was considered positive if more than 10% of the tumour cell nuclei were stained. The expression pattern of COX-2 and p53 was compared with the clinicopathological variables. RESULTS: Twenty-five patients had either synchronous or metachronous metastases and fourteen patients died during follow-up. COX-2 expression was significantly correlated only with tumour size (p=O.O49). ~53 expression was significantly correlated with TNM stage (p=O.O41), M-stage (p=O.O44), and metastasis (p=O.O29). There was no correlation between COX-2 and p53 expression (~-0.024, p=O.824). There was a statistically significant difference in the expression of p53 (p=O.O46), but not COX-2 (p=O.557), between primary and metastatic sites. Whereas the survival of patients with p53 positive tumours was significantly worse than that of patients with p53 negative tumours (p
710 NEW CELL
PROGNOSTIC CARCINOMA
Boessner
MARKERS IN THE CELL CYCLUS OF RENAL (RCC) - A FOLLOW UP OF 497 PATIENTS
P. I, Leyh H.‘, Hartung
AS CELL
A.l, Kim S.J.‘, Cho D.S.‘, Joo H.J.2, Oh D.K.‘,
of Marburg, Department
AND P53 CONVENTIONAL
R. I, Paul R.l
‘Universititsklinikum Rechts der Isar der TU Miinchen, Urologische Klinik, Munich, Germany, *Klinikum Garmisch-Partenkirchen, Urologische Klinik, GarmischPartenkirchen, Germany INTRODUCTION & OBJECTIVES: Renal cell carcinomas (RCC) have very heterogeneous tumour entities with a variety of different biological and clinical behaviours. The DNA content (ploidie) of RCC measured by flow cytometric was investigated by many study-groups in the 80’s and 90’s. But unfortunately it did not fulfil the wishes as a valuable prognostic marker. Therefore we searched for a new, prognostic more valuable factor, in the cell cycle of RCC. The aim of the study was to determine the prognostic value of the S-phase and the S+G2/M-phase in RCC cells in a retrospective study. MATERIAL & METHODS: Between 1991 and 2000 497 patients who had either a radical nephrectomy or a partial resection of the kidney because of a RCC were investigated. We quantified the fraction of the S-phase and S+G2/M-phase in the RCC and in macroscopic normal renal tissue of each patient by flow cytometric analysis. Between 2002 and 2003 a follow up concerning progression, overall survival and RCC specific survival was investigated.
711 CORRELATION BETWEEN CLINICAL OR BIOLOGICAL TUMOURS Baltoeiannis
D.l: Giannakis
CYTOGENETIC PARAMETERS
CHARACTERISTICS AND OF RENIN SECRETING
D.‘, Notsu T?, Miyagawa
I.3, Brames D.4, Sofikitis
N.’
‘Ioannina University, Department of Urology, Ioannina, Greece, *Shiritsu Hospital, Department of Surgery, Matsue, Japan, 3Tottori University, Department of Urology, Yonago, Japan, 4Cytogenetics Institute, Department of Biology, Chicago, United States INTRODUCTION & OBJECTIVES: There are no reports in the international literature concerning the chromosomal profiles of juxtaglomemlar tumours (JGT). We attempted to correlate the presence of complete or partial aneuploidies of the JGT’s chromosomes with clinical or biological parameters. MATERIAL & METHODS: JGT-tissue segments retrieved surgically from 9 patients were processed for detection of chromosomal complete and partial aneuploidies using fluorescent in situ hybridization techniques against all chromosomes and comparative genomic hybridization techniques. Preoperative clinical symptoms and signs had been recorded. Preoperative biochemical, biological, and hormonal parameters were recorded as well.
RESULTS: There was a significant difference between the median fraction of the Sphase (p
RESULTS: All the tumours had a diploid profile with complete or partial aneuploidies (monosomies or trisomies or tetrasomies) in one or more somatic chromosomes (1, 3, 8, 11, 13, 15, 16, 17, and 21). Partial aneuploidy oftheY was demonstrated in the JGT-tissue of two men. Two men and three women had partial or complete aneuploidies in chromosome 3. Participants with combined partial or complete aneuploidies in chromosomes 3 and 17 (n=4) were characterized by extremely high values of plasma renin activity (PRA) and plasma prorenin levels. The four participants with aneuploidies both in chromosome 3 and 17 were positive for malignant hypertension, thirst, pollakisuria, and severe lesions of target organs (i.e. left ventricular hypertrophy; alterations in optic fundi). The above 4 participants were the youngest within the group of the 9 men. The two men with tumours characterized by partial aneuploidies in Ychromosome (additional to other chromosomal abnormalities) demonstrated hypernatremia. Preoperatively the autonomy of tumoural renin secretion was documented through blockade of angiotensin II generation or angiotensin II receptor antagonists. High peripheral levels of histamine (that were normalized post-operatively) were observed in two men with complete/partial aneuploidies in chromosome 1.
CONCLUSIONS: The S+G2/M-Phase fraction showed the highest significance in our study concerning disease progression, overall survival and RCC-related survival and could be a useful prognostic tool in the future. Maybe the short follow-up period (2-l 1 years) is the reason for no statistical significancy in overall survival rate.
CONCLUSIONS: Combined partial or complete 17 in JGT tissue may be associated with malignant prorenin serum levels.
European
Urology
Supplements
3 (2004)
No. 2, pp. 180
aneuploidies in chromosomes 3 and forms of hypertension and very high
A.J.l, R. ’
‘University Braunschweig,
OF CELLULAR IN RENAL CELL
Hansen
W.*, Pfoertner
RETINOIC CARCINOMA
S.2, G&den
ACID-BINDING
U.2, Buer J.2, Heidenreich
709 VALUE OF PROGNOSTIC CARCINOMA
CYCLOOXYGENASE-2 INDICATORS
IN
Department of Urology, Marburg, Germany, of Cell Biology, Braunschweig, Germany
EXPRESSION RENAL
‘GBF
INTRODUCTION & OBJECTIVES: Retinoic acid (RA) is known to effect cell differentiation and proliferation and has been shown to inhibit or reverse the process of malignant transformation in several cell types. Although in vitro studies have been promising, clinical use of RA-derivates in renal cell carcinoma (RCC) did not meet first expectations. However, until today the mechanism of RA action and especially the role of the cellular retinoic acid-binding proteins (CRABP) are poorly understood. In this study. we quantified CRABP-1 mRNA expression in RCC and corresponding nonmalignant kidney tissue. Moreover, we overexpressed CRABP-1 in A498 RCC cells to disclose its potential influence on RA action in RCC. MATERIAL & METHODS: We used real-time RT-PCR methodology to investigate CRABP-1 expression in 14 RCC samples, 5 RCC cell lines (A498; CAKI-1, CAKI-2: HA-7, and LB1047) and the embryonal kidney cell line 293. Moreover, we cloned CRABP-1 to overexpress it in A498 cells via a CMV promoter. Afterwards, CRABP-1 expressing and n&e A498 cells were stimulated with at-RA and 9cis-RA; the antiproliferative effect of the RA-isotyps was evaluated in both cell types using 3H-thymidine-proliferation assays. RESULTS: The median tumour / kidney ratio for CRABP-1 expression was 0.0006 (range, 0.003.0.00002), displaying a significant downregulation of CRABP-1 in RCC. In fact, in tumour cell lines as well as in all 14 tumour samples, CRABP-1 was barely or not detectable. In contrast, CRABP-1 mRNA was expressed in embryonal 293 cells as well as in normal kidney tissue. Using proliferation assays to investigate the potential influence of CRABP-1 on the effect of RA stimulation in A498 cells, CRABP-1 expressing A498 displayed significant resistance to RA stimulation compared to nai+e A498 cells (40 and 72% at 1pM 9cis- and at-RA, respectively). CONCLUSIONS: Surprisingly, we were able to demonstrate a significant downregulation of CRABP-1 mRNA expression in all RCC cell lines as well as primary tumour samples compared to the corresponding normal kidney tissue. On the other hand, CRABP-1 overexpression in A498 RCC cells significantly inhibited RA associated antiproliferative actions. Thus, the dysregulation of CRABP-1 in RCC is probably not responsible for the limited efficiency of RA in RCC treatment.
‘Ajou University zAjou University
Choi M.K.‘,
KimYS.’
School of Medicine, Department of Urology, School of Medicine, Department of Pathology,
Suwon, South Korea, Suwon, South Korea
INTRODUCTION & OBJECTIVES: Cyclooxgenase (COX)-2 plays an important role in tumour cell proliferation, resistance to apoptosis, angiogenesis, and invasion in various cancers. However, the relationship between COX-2 expression and renal cell carcinoma (RCC) is not clear. The relationship between p53 expression and RCC is also still controversial. In this study, we investigated the expressions of COX-2 and p53 and related those to the clinicopathological variables in patients with conventional RCC. MATERIAL & METHODS: Formalin-fixed, paraffin-embedded tissue sections of conventional RCC from 92 patients who had undergone radical nephrectomy were stained immunohistochemically with specific antibodies against COX-2 and ~53. COX2 immunostaining was semi-quantitatively estimated based on proportion (percentage of positive cells) and intensity. COX-2 score was generated by multiplication of the values for the two parameters. P53 immunostaining was considered positive if more than 10% of the tumour cell nuclei were stained. The expression pattern of COX-2 and p53 was compared with the clinicopathological variables. RESULTS: Twenty-five patients had either synchronous or metachronous metastases and fourteen patients died during follow-up. COX-2 expression was significantly correlated only with tumour size (p=O.O49). ~53 expression was significantly correlated with TNM stage (p=O.O41), M-stage (p=O.O44), and metastasis (p=O.O29). There was no correlation between COX-2 and p53 expression (~-0.024, p=O.824). There was a statistically significant difference in the expression of p53 (p=O.O46), but not COX-2 (p=O.557), between primary and metastatic sites. Whereas the survival of patients with p53 positive tumours was significantly worse than that of patients with p53 negative tumours (p
710 NEW CELL
PROGNOSTIC CARCINOMA
Boessner
MARKERS IN THE CELL CYCLUS OF RENAL (RCC) - A FOLLOW UP OF 497 PATIENTS
P. I, Leyh H.‘, Hartung
AS CELL
A.l, Kim S.J.‘, Cho D.S.‘, Joo H.J.2, Oh D.K.‘,
of Marburg, Department
AND P53 CONVENTIONAL
R. I, Paul R.l
‘Universititsklinikum Rechts der Isar der TU Miinchen, Urologische Klinik, Munich, Germany, *Klinikum Garmisch-Partenkirchen, Urologische Klinik, GarmischPartenkirchen, Germany INTRODUCTION & OBJECTIVES: Renal cell carcinomas (RCC) have very heterogeneous tumour entities with a variety of different biological and clinical behaviours. The DNA content (ploidie) of RCC measured by flow cytometric was investigated by many study-groups in the 80’s and 90’s. But unfortunately it did not fulfil the wishes as a valuable prognostic marker. Therefore we searched for a new, prognostic more valuable factor, in the cell cycle of RCC. The aim of the study was to determine the prognostic value of the S-phase and the S+G2/M-phase in RCC cells in a retrospective study. MATERIAL & METHODS: Between 1991 and 2000 497 patients who had either a radical nephrectomy or a partial resection of the kidney because of a RCC were investigated. We quantified the fraction of the S-phase and S+G2/M-phase in the RCC and in macroscopic normal renal tissue of each patient by flow cytometric analysis. Between 2002 and 2003 a follow up concerning progression, overall survival and RCC specific survival was investigated.
711 CORRELATION BETWEEN CLINICAL OR BIOLOGICAL TUMOURS Baltoeiannis
D.l: Giannakis
CYTOGENETIC PARAMETERS
CHARACTERISTICS AND OF RENIN SECRETING
D.‘, Notsu T?, Miyagawa
I.3, Brames D.4, Sofikitis
N.’
‘Ioannina University, Department of Urology, Ioannina, Greece, *Shiritsu Hospital, Department of Surgery, Matsue, Japan, 3Tottori University, Department of Urology, Yonago, Japan, 4Cytogenetics Institute, Department of Biology, Chicago, United States INTRODUCTION & OBJECTIVES: There are no reports in the international literature concerning the chromosomal profiles of juxtaglomemlar tumours (JGT). We attempted to correlate the presence of complete or partial aneuploidies of the JGT’s chromosomes with clinical or biological parameters. MATERIAL & METHODS: JGT-tissue segments retrieved surgically from 9 patients were processed for detection of chromosomal complete and partial aneuploidies using fluorescent in situ hybridization techniques against all chromosomes and comparative genomic hybridization techniques. Preoperative clinical symptoms and signs had been recorded. Preoperative biochemical, biological, and hormonal parameters were recorded as well.
RESULTS: There was a significant difference between the median fraction of the Sphase (p
RESULTS: All the tumours had a diploid profile with complete or partial aneuploidies (monosomies or trisomies or tetrasomies) in one or more somatic chromosomes (1, 3, 8, 11, 13, 15, 16, 17, and 21). Partial aneuploidy oftheY was demonstrated in the JGT-tissue of two men. Two men and three women had partial or complete aneuploidies in chromosome 3. Participants with combined partial or complete aneuploidies in chromosomes 3 and 17 (n=4) were characterized by extremely high values of plasma renin activity (PRA) and plasma prorenin levels. The four participants with aneuploidies both in chromosome 3 and 17 were positive for malignant hypertension, thirst, pollakisuria, and severe lesions of target organs (i.e. left ventricular hypertrophy; alterations in optic fundi). The above 4 participants were the youngest within the group of the 9 men. The two men with tumours characterized by partial aneuploidies in Ychromosome (additional to other chromosomal abnormalities) demonstrated hypernatremia. Preoperatively the autonomy of tumoural renin secretion was documented through blockade of angiotensin II generation or angiotensin II receptor antagonists. High peripheral levels of histamine (that were normalized post-operatively) were observed in two men with complete/partial aneuploidies in chromosome 1.
CONCLUSIONS: The S+G2/M-Phase fraction showed the highest significance in our study concerning disease progression, overall survival and RCC-related survival and could be a useful prognostic tool in the future. Maybe the short follow-up period (2-l 1 years) is the reason for no statistical significancy in overall survival rate.
CONCLUSIONS: Combined partial or complete 17 in JGT tissue may be associated with malignant prorenin serum levels.
European
Urology
Supplements
3 (2004)
No. 2, pp. 180
aneuploidies in chromosomes 3 and forms of hypertension and very high