Bicalutamide 80 mg combined with a luteinizing hormone-releasing hormone agonist (LHRH-A) versus LHRH-A monotherapy in advanced prostate cancer: findings from a phase III randomized, double-blind, multicenter trial in Japanese patients.

To compare combination therapy with bicalutamide 80 mg and a luteinizing hormone-releasing hormone agonist (LHRH-A) versus LHRH-A alone in Japanese men with untreated advanced prostate cancer. A total of 205 patients with stage C/D prostate cancer were randomized to either LHRH-A+once-daily oral bicalutamide 80 mg or placebo. Primary study variables have been reported previously. Secondary variables included: time to achieve prostate-specific antigen < or = 4 ng/ml, time-to-treatment failure (TTTF), time-to-disease progression (TTP), overall survival (OS), adverse events and adverse drug reactions. Following combination therapy with bicalutamide 80 mg, there were significant (P<0.001) advantages over LHRH-A alone in terms of TTTF and TTP, but the difference in the interim OS was not statistically significant. First-line combination therapy with bicalutamide 80 mg in Japanese patients with advanced prostate cancer offers significant benefits over LHRH-A alone, with respect to TTTF and TTP. Follow-up for OS continues.

Increase in hepatocyte growth factor receptor tyrosine kinase activity in renal carcinoma cells is associated with increased motility partly through phosphoinositide 3-kinase activation.

Dysregulated cell motility is one of the major characteristics of invasion and metastatic potentials of malignant tumor cells. Here, we examined the hepatocyte growth factor (HGF)-induced cell motility of two human renal carcinoma cell lines, ACHN and VMRC-RCW. Scattering and migration was induced in ACHN in an HGF-dependent manner, whereas they were maintained in VMRC-RCW even in the absence of HGF. In VMRC-RCW, HGF receptor (HGFR) tyrosine kinase was constitutively active, and sequence analysis showed N375S, A1209G and V1290L mutations. However, transfection experiments using porcine aortic endothelial (PAE) cells demonstrated that no single mutation or combination of two or three mutations caused HGF-independent constitutive activation. Conversely, the expressed amount of receptor protein had a pivotal role in the basal kinase activity. With respect to downstream signaling molecules of HGFR in ACHN or VMRC-RCW, the Ras-MAPK pathway was downregulated, whereas phosphoinositide 3-kinase (PI3-kinase) was not further activated by HGF-treatment in VMRC-RCW cells. The PI3-kinase inhibitors, wortmannin and LY294002 strongly inhibited spontaneous migration of VMRC-RCW. One transfected PAE cell line with massive overexpression of HGFR demonstrated scattered morphology and increased PI3-kinase activity in association with increased motility, which was partially inhibited by LY294002. Taken together, our results indicate that the overexpression of HGFR causes increase in cellular motility and PI3-kinase shows the important contribution on the increased motility of renal carcinoma cells.

Expression profile of prostate-specific antigen messenger RNA assessed by in situ hybridization is a novel prognostic marker for patients with untreated prostate cancer.

The present study was undertaken to define the relationship between histological grade (Gleason grade) and prostate-specific antigen (PSA) mRNA expression and to evaluate the level of PSA mRNA expression as a possible prognostic marker for untreated prostate cancers. The primary grade areas of 104 prostatic biopsy specimens were analyzed for the expression of PSA mRNA and its protein by nonradioactive in situ hybridization and immunohistochemistry, respectively. A multivariate survival analysis was performed to examine the correlation between PSA mRNA expression and several clinicopathological parameters, e.g., the immunostaining level of PSA protein in biopsy specimens. The percentage of specimens positive for PSA mRNA increased significantly with advanced histological grade. Image analysis of the signal intensity for PSA mRNA showed a significant correlation between the signal intensity in both primary and secondary grade areas of each specimen and the histological grade (P < 0.0001). Only 26.0% of specimens positive for PSA protein were also positive for PSA mRNA (and vice versa, 6.7%). Other tumors were either positive for both (66.3%) or negative for both (1.0%). When the Cox's proportional hazards regression model was used to analyze cancer-specific survival, untreated patients with higher levels of PSA mRNA expression in the higher grade (representing higher grade of either primary or secondary grade) area of tumors were at high risk for cancer-related death (P = 0.017). Furthermore, in cancer-specific survival curves based on PSA mRNA expression status, patients with high levels of PSA mRNA expression in the higher grade area of tumors had a significantly poorer prognosis (P = 0.001), compared with those with tumors expressing low levels of PSA mRNA. Our results suggested that analysis of PSA mRNA expression in specific areas in biopsy specimens of patients with untreated prostate cancer may provide a good assessment of prognosis of prostate cancers.

All-trans retinoic acid enhances gap junctional intercellular communication among renal epithelial cells in vitro treated with renal carcinogens.

Epidemiological and clinical studies imply that retinoids have a chemopreventative action against cancer and can suppress the growth of cancer cells. The regulation of connexin (Cx) expression by retinoids varies among tissues and organs. In this study, we investigated whether all-trans retinoic acid (ATRA) upregulates gap junctional intercellular communication (GJIC) in renal epithelial cells exposed to renal carcinogens. Madin Darby canine kidney (MDCK) cells were incubated with ATRA for 3 days, then briefly exposed to 12-O-tetradecanoyl-phorbol-13-acetate (TPA) or renal carcinogens potassium bromate (KBrO3) and dimethylnitrosamine (DMN). ATRA increased the expression of connexin 43 mRNA and protein without affecting Cx 43 phosphorylation and prevented inadequate Cx 43 localisation caused by TPA/KBrO3 or DMN. Consequently, ATRA prevented the disruption of GJIC in MDCK cells. These data suggest that ATRA enhanced GJIC by upregulating Cx 43 expression and that ATRA might be useful for prevention of renal cell carcinoma.

The expression of platelet-derived endothelial cell growth factor in human bladder cancer.

Platelet-derived endothelial cell growth factor (PD-ECGF) is an angiogenic factor and in some studies PD-ECGF/dTdRPase expression levels in several kind of cancers were higher than in their surrounding normal tissues. In this study, we evaluated PD-ECGF/dTdRPase expression in bladder cancer by an immunohistological method and determined whether it correlated with tumor stage, grade and recurrence. PD-ECGF/dTdRPase expression was correlated with tumor grade and stage. Furthermore, among the superficial tumors, PD-ECGF/dTdRPase expression was correlated with a recurrence-free rate and thus it might be a prognostic factor for the recurrence of superficial bladder cancer.

Changes in the gap junctional intercellular communication in renal tubular epithelial cells in vitro treated with renal carcinogens.

Gap junctional intercellular communications (GJIC) are known as the channels for the direct transfer of cytoplasmic molecules between neighboring cells and are lost during transformation of normal cells. To study the function and the molecular mechanism for the loss of GJIC, the effects of dimethylnitrosamine, KBrO3 and FeSO4 x 7H2O, which are known as chemical tumor promoters of the kidney on the GJIC function and the expression of connexin 43 of Madin-Darby canine kidney (MDCK) epithelial cells, were examined. These tumor promoters inhibited the GJIC in MDCK cells. The expression of connexin 43 mRNA and connexin 43 protein was not altered by these treatments, whereas immunocytochemical study revealed that the distribution of connexin 43 protein was changed from the cell surface to the cytoplasma. These data suggest that blockage of GJIC in MDCK cells treated with renal carcinogens support the hypothesis that loss of GJIC might be important in renal carcinogenesis.

Effects of low concentration of vinblastine on the anchorage-independent growth and in vitro invasion of human renal carcinoma cell lines.

A study was conducted on the effect of vinblastine (VBL), an anti-mitotic drug that is commonly employed in the treatment of human renal cell carcinoma. When VBL was added to serum-free cultures of the ACHN and NT cell lines (both lines are of human renal carcinoma origin), a concentration of 1 microgram/ml resulted in death of most of the cells of both cell types. However, at a concentration of 10 ng/ml or less, although the cells detached from the culture dish, many viable cells were observed. In addition, in an in vitro invasion assay, the invasiveness of these detached cells was demonstrated to be accelerated in comparison with the parent monolayed cells. This increase of invasion was observed in the treatment of TN-16 which is known to have a metaphase-arresting effect, not to have an anti-cancer effect. When detached cells by VBL were inoculated into soft agar, their colony-forming ability was clearly increased in comparison with the parent cells or TN-16 treated cells. These results indicate that low concentration of VBL appears to increase the malignant potential of human renal carcinoma cells in culture.

Preparation of monoclonal antibodies against N-(gamma-maleimidobutyryloxy)succinimide (GMBS)-conjugated acetylspermine, and development of an enzyme-linked immunosorbent assay (ELISA) for N1,N12-diacetylspermine.

We have developed three mouse monoclonal antibodies (mAb) of types IgG1 and IgG2b, i.e. anti-acetylspermine (Ac-Spm)-1 and 2 (ACSPM-1 and 2), and anti-acetylspermine (Ac-Spm)-3 (ACSPM-3), respectively, against Ac-Spm conjugated to bovine serum albumin via a heterobifunctional cross-linker, N-(gamma-maleimidobutyryloxy)succinimide (GMBS). Among these mAbs, ACSPM-2 was the most useful for the development of an enzyme-linked immunosorbent assay (ELISA) for acetylpolyamines (Ac-PAs) with glutaraldehyde (GA)-conjugated N1,N12-diacetylspermine (2Ac-Spm) or acetylspermine (Ac-Spm) as the solid phase antigen. However, GMBS-conjugated Ac-Spm did not behave as a solid phase antigen in the competitive ELISA. The ELISA is based on the principle of competition between an analyte and the conjugated antigen for the mAb, followed by immunoreaction with biotinylated anti-mouse immunoglobulin and horseradish peroxidase-streptavidin. The ACSPM-2 mAb reacted with 2Ac-Spm to the highest degree, followed by Ac-Spm, N1-acetylspermidine (N1-Ac-Spd), N8,N8-diacetylspermidine (2Ac-Spd), and spermine (Spm), the EC50 values being 0.06, 0.25, 7.0, 10, and 60 microM, respectively, but exhibited almost no cross-reaction with other polyamine-related compounds or amino acids. The method was used to determine the urinary Ac-PA levels in healthy subjects, the average value of 0.36 microg of 2Ac-Spm/g creatinine (n = 16) being obtained. The ACSPM-2 ELISA for 2Ac-Spm, which was the PA most relevant to the analysis of human urine among the five PA analogs mentioned above, might have potential for elucidation of the correlation of urinary 2Ac-Spm levels in cancers.

Determination of urinary acetylpolyamines by a monoclonal antibody-based enzyme-linked immunosorbent assay (ELISA).

A monoclonal antibody (mAb), ASPM-2, produced against N-(gamma-maleimidobutyryloxy)-succinimide (GMBS)-conjugated polyamine spermine [Spm; Fujiwara et al. (1994) Histochemistry 102, 397-404] was used for the development of an enzyme-linked immunosorbent assay (ELISA) for acetylpolyamines (Ac-PAs) in human urine. The ELISA is based on the principle of competition between an analyte and Spm-glutaraldehyde-bovine serum albumin conjugate-coated polystyrene microtiter wells for the mAb, followed by immunoreaction with biotinylated anti-mouse immunoglobulin and horseradish peroxidase-streptavidin. The ASPM-2 mAb showed strong immunoreaction with N1,N12-diacetylspermine (2Ac-Spm), N-monoacetylspermine (Ac-Spm), and N1-acetylspermidine (N1-Ac-Spd), the EC50 values being 29, 50, and 51 microM, respectively, but no cross-reaction with other PA-related compounds or amino acids. The method was used to measure urinary Ac-PA levels in healthy subjects and cancer patients, without pretreatment of the specimens, mean concentrations of 3.25 and 2.80 mumol per 24-h urine, respectively (as N1-Ac-Spd), being found. The ASPM-2 ELISA for N1-Ac-Spd, which is the PA most relevant to the analysis of human urine among the three Ac-PAs mentioned above, is specific and accurate, and can easily be used to analyze large numbers of specimens in parallel. It should thus have potential for studying the relationship between urinary N1-Ac-Spd levels and cancer.

Outcome of surgery in cystic renal cell carcinoma.

OBJECTIVES: To review cases of cystic renal cell carcinoma treated surgically at our institution and define their clinical and histopathologic features. METHODS: Between 1986 and 1998, 21 patients with cystic renal cell carcinoma were treated surgically. Cystic renal cell carcinoma was categorized using Hartman's classification. RESULTS: Histopathologic examination demonstrated cystic necrosis in 11 patients, multilocular cystic renal cell carcinoma in 9, and unilocular cystic renal cell carcinoma in 1 patient. Tumors were incidentally found during an evaluation of unrelated disease or a general health checkup in 14 patients (67%). The mean tumor size was 5.6 cm (range 0.5 to 12) for cystic necrosis and 5.4 cm (range 2 to 9) for multilocular cystic renal cell carcinoma. All 9 cases of multilocular cystic renal cell carcinoma were of the clear cell type and tumor grade 1. The mean follow-up period was 65 months (range 9 to 141). The 5-year disease-specific survival rates for multilocular cystic renal cell carcinoma and cystic necrosis were 100% and 80%, respectively. CONCLUSIONS: The prognosis for patients with cystic renal cell carcinoma is better than that for patients with solid tumors. In particular, the prognosis of multilocular cystic renal cell carcinoma is excellent. Multilocular cystic renal cell carcinoma represents a distinct subtype of renal cell carcinoma that can be completely cured by surgery.

Disruption of gap junctional intercellular communication in human renal cancer cell lines.

OBJECTIVES: Gap junctional intercellular communication (GJIC) is believed to play an important role in the maintenance of cell homeostasis, and its disruption may be associated with carcinogenesis. However, GJIC has not been detected in many human cancers. We therefore studied the regulation of GJIC in human renal cancer cell lines. METHODS: We examined the human renal cancer cell lines, ACHN and NT, as well as Madin-Darby canine kidney (MDCK) cells as a positive control, using GJIC assays, Northern blotting to detect connexin 43 mRNA, immunofluorescent staining, and Western blotting of connexin 43 protein. RESULTS: GJIC of ACHN and NT was completely blocked. In ACHN cells, connexin 43 mRNA was not altered. However, connexin 43 protein was aberrantly localized and phosphorylated connexin 43 protein had disappeared. Both connexin 43 protein and its mRNA were undetectable in NT cells. CONCLUSIONS: GJIC in human renal cancer cell lines is impaired and various pathways may inhibit this mechanism in renal cancer. We believe that connexin plays an important role in renal carcinogenesis.

Chemo-endocrine therapy for prostate cancer with bone metastasis. Nagasaki Prostate Cancer Research Group.

We analyzed the clinical effects of initial chemoendocrine therapy on 31 prostate cancer patients with bone metastasis. These patients had been newly diagnosed between 1983 and 1991 and had received no previous therapy. As endocrine therapy, the patients received 1 mg ethynylestradiol daily with or without orchiectomy. In addition, they received three courses of chemotherapy consisting of 20 mg/m2 cisplatin given on days 1, 3, and 5 and 20 mg/m2 Adriamycin or 40 mg/m2 epirubicin given on day 5. Subsequently, for maintenance therapy, the patients received 1 mg ethynylestradiol and 150 mg 5-fluorouracil [or 300 mg tegafur plus uracil (UFT)] daily. Patients given our regimen of chemoendocrine therapy had a significantly better prognosis than did the controls treated with endocrine therapy alone (P = 0.05), although treatment was not randomized. The cause-specific survival rates at 5 years for the chemoendocrine-therapy patients and the control group were 65.4% and 37.4%, respectively. A multivariate analysis of possible prognostic factors, i.e., age, histological grade, prostatic acid phosphatase, tumor-related pain, the extent of disease (EOD) on bone scan, and the type of initial treatment, confirmed that the initial treatment (P = 0.03) and the EOD grade (P = 0.05) had a significant effect on survival. On the basis of these results, it is necessary to carry out a randomized trial to compare our chemoendocrine regimen with endocrine therapy alone in untreated patients with advanced prostate cancer.

Subcutaneous administration of interferon alpha and gamma in patients with metastatic renal cell carcinoma.

BACKGROUND: Renal cell carcinoma (RCC) is relatively resistant to both chemotherapy and radiotherapy. Response and survival of treatment with Interferon-alpha (IFN-alpha) and Interferon-gamma (IFN-gamma) were evaluated in patients with metastatic RCC. PATIENTS AND METHODS: Thirty-one patients with confirmed RCC were included in this study. Fifteen of 31 patients received injection of IFN-alpha and IFN-gamma three times a week. IFN-gamma was infused subcutaneously by microinfusion pump. Sixteen received IFN-alpha alone more than three times a week. RESULTS: The overall response rate was 20.0% in the IFN-alpha and IFN-gamma group, and 12.5% in the IFN-alpha alone group. Long lasting stabilization of the disease (more than; 12 months) was seen in 92.3% of CR, PR or SD in the IFN-alpha and IFN-gamma group, as compared with 71.4% in the IFN-alpha alone group. Both groups differed significantly in survival rate from the first treatment with IFN (p < 0.05). CONCLUSIONS: A long lasting stabilization of the disease can be expected in patients who were treated with our regimen of IFN-alpha and IFN-gamma.

TAC-101, a novel retinobenzoic-acid derivative, enhances gap junctional intercellular communication among renal epithelial cells treated with renal carcinogens.

[4-3,5-Bis(trimethylsilyl)benzamido] benzoic acido] (TAC-101), which exhibits an anti-tumor effect, can bind to retinoic acid receptors (RARs). It has retinoid-like properties, such as chemopreventive action against cancer cells. The up-regulation of connexin (Cx) expression by retinoids is well known in various epithelial cells. In this study, we investigated whether TAC-101 up-regulates gap junctional intercellular communication (GJIC) in renal epithelial cells exposed to the renal carcinogens. Madin Darby canine kidney (MDCK) cells were incubated with TAC-101 for 3 days, then briefly exposed to renal carcinogens potassium bromate (KBrO3) or dimethylnitrosamine (DMN). TAC-101 increased the expression of connexin 43 protein without affecting Cx43 phosphorylation and prevented inadequate Cx43 localisation caused by KBrO3 or DMN. Consequently, TAC-101 prevented the disruption of GJIC in MDCK cells. These data suggested that TAC-101 enhanced GJIC by up-regulating Cx43 expression and that TAC-101 might be useful for the prevention of renal cell carcinoma.

Renal cell carcinoma metastatic to the skin.

BACKGROUND: Cutaneous metastases from renal cell carcinoma are rarely diagnosed during life. We reviewed the cases of our institute and from the Japanese literature and analyzed them to define the clinical features. PATIENTS AND METHODS: Six patients with cutaneous metastases from renal cell carcinoma were treated in our institute. Clinical data and follow-up information of these patients were reviewed. RESULTS: The skin metastatic lesions were found before the renal lesion was discovered in one case and they were found after the diagnosis had been established in 5. Four patients presented with solitary cutaneous metastasis at the time of diagnosis of the skin metastatic lesion. Five patients had other sites of metastases such as the lung, liver and bone at the time of diagnosis of cutaneous metastases. Five patients died of renal cell carcinoma at mean 23.8 months after presenting with cutaneous metastases. CONCLUSION: Our 6 cases bring the total reported in the Japanese literature to 75 cases. The results of these 75 cases revealed that the duration from the time of diagnosis of renal cell carcinoma to detection of the cutaneous metastasis was relatively long but the prognosis of patients with these metastatic skin lesions was poor. Cutaneous metastases from renal cell carcinoma were regarded as a late manifestation of the disease.

Breast cancer metastatic to the kidney.

We report a case of renal tumor secondary to a breast cancer occurring 16 years after radical mastectomy. This is the sixth report case of renal metastasis from breast cancer of a 51-year-old woman. Percutaneous biopsy of the renal tumor confirmed the diagnosis during the follow-up. The patient was treated with chemotherapy and is alive 8 months after diagnosis. Previously, cases like our case showed long interval from mastectomy to diagnosis of metastasis.

The role of surgery in renal cell carcinoma with solitary metachronous metastasis to contralateral adrenal gland.

We report a case of renal cell carcinoma with solitary metachronous metastasis to contralateral adrenal gland occurring 9 months after radical nephrectomy. The patient was treated with a contralateral adrenalectomy and is alive for 87 months. The literature was reviewed and 5-year survival of solitary metachronous metastasis to contralateral adrenal gland was 60%. Follow-up duration of our case was the longest in the literature. It is suggested that the solitary contralateral adrenal gland metastasis of renal cell carcinoma should be resected since there is no effective treatment of metastatic renal cell carcinoma. Good prognosis may be then and the good be expected.

Kidney transplantation from non-heart-beating donors: analysis of organ procurement and outcome.

INTRODUCTION: Most donors in Japan have been non-heart-beating donors (NHBD), so-called "marginal donors." In Western countries kidney transplants from NHBD have also been increasing. We analyzed 120 kidneys harvested from NHBD with regard to organ procurement, renal function, graft survival, and the donor factors that affected graft survival. METHODS: Donors were moved into the operating room after cardiac arrest. A double-balloon catheter was inserted into the abdominal aorta via laparotomy. In situ cooling by Euro-Collins solution was started at 500 mL/min. We did not performed cannulation into the femoral artery or vein prior to cardiac arrest. RESULTS: Warm ischemia time (WIT) was 18.6 minutes. Among 108 kidneys (90%) used for transplantation, 102 kidneys functioned. There were no cases of bilateral nonfunctioning kidneys. The delayed graft function (DGF) rate was 86%; however, the death-censored graft survival was 80.0% at 5 years and 62.9% at 10 years. Kidneys implanted after more than 24 hours of total ischemia time required a significantly longer period of hemodialysis. Donor risk factors that affected graft survival included WIT >/= 20 minutes, donor age >/= 50 years, and serum creatinine level at admission > 1.0 mg/dL. CONCLUSIONS: Organ procurement without cannulation prior to cardiac arrest entailed a long WIT and a high DGF rate. However, the graft survival was good. It has been necessary to use grafts from NHBD despite the inherent risk factors. It is important to reduce kidney damage both at the organ procurement and during the posttransplant management.

Analysis of survival of prostate cancer patients in Japan and the U.S.A.

The prognosis of U.S.A. patients with prostate cancer based on reports from the American College of Surgeons and Roswell Park Cancer Institute was compared with that of Japanese in Nagasaki and Gunma. There was more than a 10% difference in survival rate between the two countries. Several clinical parameters were analyzed to determine the basis for this difference. In general, stage distribution, metastatic sites, bone pain and alkaline phosphatase in the USA patients with Stage D disease were more severe than in the Japanese patients. Moreover the prostate cancer death rate in the USA was higher than that in Japan. However, only the distribution of pathological grade in the USA was better than that in Japan. From these results, it is difficult to conclude that prostate cancer in the USA is more aggressive than in Japan. The possibility of selection bias in these studies is discussed.

Segmental stenosis of ureter: a late complication of intra-arterial chemotherapy for bladder cancer.

A 54-year-old Japanese male was treated with a single shot of cisplatin-phosphatidylcholine-lipiodol (CPL) suspension due to bladder tumour (stage T2N0M0). Seven months later, a right lower ureteral stenosis developed. The possible cause of ureteral stenosis due to intra-arterial chemotherapy is discussed.