[A case of dedifferentiated liposarcoma of the spermatic cord].

A 61-year-old man was referred to our department because of painless and stony hard mass beside the left testis. Serum levels of lactate dehydrogenase, alpha-fetoprotein and human chorionic gonadotropin were within normal ranges. The ultrasonography of the mass showed almost homogenous and relatively low intensity echogram. The mass which derived from the left spermatic cord and was partially surrounded by fat-like soft and yellow tissue, was removed with the left testis by usual orchiectomy. Histopathological diagnosis was liposarcoma, whose subtype was dedifferentiated type derived from well differentiated type. Postoperatively, a para-aortic mass, which resembled lymphnode metastasis, was pointed by computed tomographic scan and was removed surgically. However, it was histopathologically diagnosed as neurogenicganglioma irrelevant to liposarcoma. He has been free of disease for about 1 year without any adjuvant therapy.

[A case of primary mucinous adenocarcinoma of the scrotal contents].

A 48-year-old man consulted our hospital complaining of painless swelling of the left scrotal contents that had gradually increased for 5 years. Serum LDH, alpha-fetoprotein and HCG were within normal ranges. The ultrasonography showed heterogeneous echogram including high-echogenic spots and relatively a low-echogenic hydrocele. Left high orchiectomy was performed and the removed tissue was 24 x 16 x 15 cm in size, which had large cystic cavity filled with cloudy and deep-green mucin. Pathological diagnosis was mucinous adenocarcinoma. Postoperatively analyzed serum CEA, CA19-9 and PSA were within normal ranges. Systemic X-ray examinations, such as lung, abdominal and pelvic CT scan, upper GI series and barium enema, did not show any abnormal SOL suggesting carcinoma. Therefore, we diagnosed this case as a primary adenocarcinoma in the scrotal contents. The patient has been observed without any adjuvant therapy since operation, but no signs of recurrence have been identified for one year and six months.

Usefulness of F-18 FDG-PET in a long-term hemodialysis patient with renal cell carcinoma and pheochromocytoma.

A patient who had been on long-term hemodialysis (HD) was diagnosed as having renal cell carcinoma (RCC) and pheochromocytoma. Abdominal computed tomography scanning demonstrated a right renal mass and a right adrenal mass, whereas positron emission tomography (PET) using F-18 fluorodeoxyglucose (FDG) revealed increased accumulation in both the renal and adrenal masses. FDG-PET is useful for detecting RCC in HD patients because FDG is not excreted in the urine, but it is difficult to distinguish pheochromocytoma from an adrenal metastasis by this imaging method.

Palliative treatment of bone metastases in hormone-refractory prostate cancer: effects of pamidronate on the carboxyterminal telopeptide of type-I collagen level in patients with increasing prostate-specific antigen levels.

PURPOSE: Bisphosphonates have been reported to be effective in reducing bone pain and skeletal-related events associated with bone metastases in hormone-refractory prostate cancer (HRPC). However, whether bone resorption is reduced primarily by these particular drugs is difficult to evaluate because patients with HRPC are usually treated with secondary or tertiary hormonal manipulations including second-line antiandrogens, high-dose diethylstilbestrol, or low-dose dexamethasone therapies, some of which may also be effective. Thus, we assessed changes in the level of the carboxyterminal telopeptide of type-I collagen (ICTP), a bone resorption marker, before and after pamidronate administration in HRPC patients with increasing prostate-specific antigen (PSA) levels. PATIENTS AND METHODS: Twenty-one HRPC patients with bone metastases and increasing PSA levels were intravenously treated with pamidronate at a dose of 30 mg either every 2 or every 4 weeks. Pamidronate administration was started immediately after confirmation of three consecutive increases in the PSA level. RESULTS: In 14 patients (67%), the ICTP levels decreased after the administration of pamidronate, despite increasing PSA levels. In 7 of these cases, the ICTP levels were lower than those recorded for 6 months or longer before the start of pamidronate administration. The characteristics of the responders were compared with those of the non-responders. CONCLUSION: In 67% of the HRPC patients with increasing PSA levels, pamidronate reduced the accelerated turnover of bone metabolism caused by metastases of prostate cancer.

Significance of target cell infection and natural killer cells in the anti-tumor effects of bacillus Calmette-Guerin in murine bladder cancer.

Although intravesical instillation of bacillus Calmette-Guerin (BCG) is a clinically well-recognized therapy for bladder carcinoma in situ and recurrence prophylaxis, these mechanisms have not been fully understood. We studied the effects of BCG infection (Connaught strain) on target cancer cells and host immune systems in murine bladder cancer. The bladder cancer cell line, MB49, was used in C57/BL6 mice in vivo and in vitro. In vitro cytotoxicities against the cancer cell line were measured by 24-h 51Cr release assay. For effector cells, spleen mononuclear cells were obtained from mice injected intraperitoneally with BCG or BCG-infected irradiated MB49 cells. Although BCG infection of cancer cells did not affect the proliferation speed in vitro, the mice injected subcutaneously with BCG-infected MB49 cells survived significantly longer than those given untreated cancer cells. The mice surviving without tumor growth after injection of BCG-infected cancer cells could not reject a second injection of intact MB49 cells. In vitro cytotoxicity was enhanced by BCG infection of target cancer cells, but not by immunizing the mice with BCG from which effector cells were obtained. Moreover, cytotoxicity disappeared by depleting natural killer (NK) cells from effector cells. Although in vitro cytotoxicity was increased by immunizing the mice with BCG-infected irradiated MB49 cells, survival did not improve in these mice. These results suggest that a major part of BCG's anti-tumor effects can be attributed to the elimination of BCG-infected cancer cells by NK cells.

Comparative antitumor activity of 5-fluorouracil and 5'-deoxy-5-fluorouridine in combination with interferon-alpha in renal cell carcinoma cell lines.

OBJECTIVES: Although interferon-alpha (IFNalpha) and interleukin-2 are used in the treatment of advanced renal cell carcinoma (RCC), the rate of efficacy is about 15% and not satisfactory. Therefore, a more effective treatment is being investigated. In this study, we examined the combined effects of IFNalpha and 5-fluorouracil (5-FU) as well as 5-deoxy-5-fluorouridine (5'-DFUR), a produrg of 5-FU, in vitro. MATERIALS AND METHODS: Four RCC cell lines (OCUU1, OCUU3, OCUU4, OCUU5) were established at our laboratory from RCC patients. OS-RC-2, RCC10RGB, TUHR14TKB, TUHR4TKB, A498 and Caki-1 were obtained from a commercial source. The sensitivity of the 10 RCC cell lines to 5-FU and 5'-DFUR was evaluated using MTT assay. The IC50 value for the cytostatics was expressed as the concentration at which growth was inhibited by 50% as compared with the control value. Thymidine phosphorylase (TP), the enzyme that converts 5'-DFUR into 5-FU and 5-FU into FdUMP, was estimated by ELISA. RESULTS: When the 10 RCC cell lines were divided into the low TP expression group and high TP expression group at 2.0 U/ml protein, TP expression was not enhanced by IFNalpha in all 4 cell lines in the low TP expression group. Antitumor effects were not enhanced by IFNalpha in 3 out of 4 cell lines for 5-FU and in all 4 cell lines for 5'-DFUR. On the other hand, in the high TP expression group, TP expression was enhanced by IFNalpha in 5 out of 6 cell lines, and antitumor effects were enhanced by IFNalpha in 5 out of 6 cell lines for 5-FU and in all 6 cell lines for 5'-DFUR. In addition, there was a significant correlation between TP expression and sensitivity to 5-FU and 5'-DFUR in all RCC cell lines. CONCLUSIONS: These results suggested that TP may be useful as a predictive factor in combination therapy with IFNalpha and 5-FU or 5'-DFUR, which may be a promising treatment for advanced RCC.

Chondrosarcoma of the urinary bladder and establishment of a human chondrosarcoma cell line (OCUU-6).

Chondrosarcoma is a very rare tumor of the urinary bladder, with only 4 cases reported to date. In this study, we report on a case of a 73-year-old male who presented bladder mass and right hydroureteronephrosis. Radical cystectomy, right nephrectomy and left ureterocutaneoustomy were performed, and histological study disclosed chondrosarcoma of the urinary bladder. As reported in other cases, the tumor was highly aggressive with a short clinical course, and the patient died of carcinomatous pleuritis at one month after surgery. Subsequently, we successfully established a human chondrosarcoma cell line (OCUU-6) from the pleural effusion of the patient.

Antitumor effects of lipoxygenase inhibitors on murine bladder cancer cell line (MBT-2).

Arachidonic acid (AA) metabolites derived from both the lipoxygenase (LOX) and cyclooxygenase (COX) pathways transduce a variety of signals related to cell growth. Selective blockade of the different metabolic pathways of AA (using a general LOX inhibitor NDGA, a 5-LOX inhibitor AA861, a 12-LOX inhibitor baicalein and a general COX inhibitor ibuprofen) revealed that murine bladder cancer cell line (MBT-2) cell proliferation was inhibited by the LOX inhibitors concentration-dependently, but not by the COX inhibitor. Among the LOX inhibitors, baicalein showed the strongest inhibition and induced apoptosis of MBT-2. Proliferation of MBT-2 was also significantly inhibited by 12-LOX antisense oligonucleotides. In an in vivo experiment, the antitumor effects of baicalein administration on C3H/HeN mice implanted with MBT-2 were recognized. These results suggested that LOX inhibition may be significant in the treatment of bladder cancer.

Chondrosarcoma of the urinary bladder and establishment of a human chondrosarcoma cell line (OCUU-6).

Chondrosarcoma is a very rare tumor of the urinary bladder, with only 4 cases reported to date. In this study, we report on a case of a 73-year-old male who presented bladder mass and right hydroureteronephrosis. Radical cystectomy, right nephrectomy and left ureterocutaneoustomy were performed, and histological study disclosed chondrosarcoma of the urinary bladder. As reported in other cases, the tumor was highly aggressive with a short clinical course, and the patient died of carcinomatous pleuritis at one month after surgery. Subsequently, we successfully established a human chondrosarcoma cell line (OCUU-6) from the pleural effusion of the patient.

Role of tumor-associated macrophages in renal cell carcinoma.

We investigated the biologic meaning of tumor-associated macrophages (TAM) in renal cell carcinoma (RCC). The study group comprised of 83 patients with RCC. TAM was isolated by plastic adherence following enzymatic digestion of surgically removed tumor tissues. In some of the patients, monocytes were also isolated from peripheral blood mononuclear cells by plastic adherence. When TAM and monocytes were compared in the same patients, TAM produced interleukin 6 (IL-6), tumor necrosis factor alpha (TNFalpha) and interleukin 1beta (IL-1beta) without lipopolysaccharide (LPS) stimulation, while monocytes hardly produced IL-6, TNFalpha and IL-1beta without LPS stimulation. However, with LPS stimulation, monocytes produced more IL-6, TNFalpha and IL-1beta than TAM. In stage T1 RCC patients, there was a significant positive correlation between TNFalpha production of TAM and tumor size. In order to investigate the effects of TAM on cancer cells, TAM was co-cultured with A498, K562 and in some cases, with short-term established RCC lines for 96 h. As a result, TAM largely enhanced cell proliferation. These results suggested that TAM may play an important role in certain steps of tumor progression.

Effects of tumor necrosis factor alpha in renal cell carcinoma.

Tumor necrosis factor alpha (TNFalpha) and TNFalpha receptor mRNA expression, the effects of TNFalpha on DNA synthesis and cell proliferation as well as its effects on interleukin-6 (IL-6) production and expression in renal cell carcinoma (RCC) were studied using RCC cell lines. The effects of TNFalpha on DNA synthesis and IL-6 production were also studied using short-term established RCC cell lines. A total of 8 cell lines, 4 RCC cell lines (RC-2, RGB, 14T, 4T) and 4 cell lines established at our laboratory (OCUU-1, 2, 4, 5), as well as 10 short-term established RCC cell lines were used. When TNFalpha and TNFalpha receptor mRNA expression was examined by RT-PCR, p55 TNF receptor mRNA expression was observed while TNFalpha and p75 TNF receptor mRNA expression was not observed in all cell lines. When the effects of TNFalpha on DNA synthesis were studied by [3H]-thymidine incorporation assay, DNA synthesis was induced in RGB, 14T, 4T, OCUU-2 and OCUU-4 by adding 10 and 100 pg/ml of TNFalpha while it was not induced in RC-2 and OCUU-5 at all concentrations. When its effects on cell proliferation were evaluated by MTT assay, cell proliferation was observed in RGB, 14T and 4T at TNFalpha concentration of 10 pg/ml and in RGB, 14T, 4T, OCUU-4 and OCUU-5 at TNFalpha concentration of 100 pg/ml. However, cell proliferation was not detected in RC-2 and OCCU-2 at any concentration. When the effects of TNFalpha on IL-6 production were studied by ELISA, IL-6 production was induced in RC-2, RGB, 14T, OCUU-1, OCUU-2 and OCUU-5 while not in 4T and OCUU-4. When its effects on IL-6 expression were examined by Northern blot analysis, the results were similar to those obtained by ELISA. As for the 10 short-term established RCC cell lines, DNA synthesis and IL-6 production were induced with the addition of TNFalpha. These results suggested that TNFalpha induced cell proliferation in RCC.

Norepinephrine activates P44 and P42 MAPK in human prostate stromal and smooth muscle cells but not in epithelial cells.

BACKGROUND: In vascular smooth muscle cells, alpha1-adrenergic stimulation increases DNA synthesis and cell proliferation via activation of p44/42 (ERK1/2) MAPK. We examined whether norepinephrine (NE) activates MAPK and stimulates the proliferation of prostatic epithelial and non-epithelial cells. METHODS: Human prostatic epithelial cells, stromal cells, and smooth muscle cells were purchased from BioWhittaker (Walkersville, MD). After reaching a semi-confluent condition, the cells were cultured in RPMI-1640 without serum for 1 day. At 10 min after adding NE (10(-6) or 10(-7) M) to the medium, the cells were collected. Cell lysate was analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) followed by Western blot using anti-phospho-p44/42 and anti-p44/42 antibodies. The activation of p44/42 was estimated by the ratio of phospho-p44/42 to total p44/42. Cell proliferation was evaluated by (3)H-thymidine uptake assay. After reaching a semi-confluent condition, the cells were cultured in RPMI-1640 containing 0.5% FCS with or without NE (10(-6) or 10(-7) M) for 16 hr followed by a (3)H-thymidine uptake period (24 hr). RESULTS: P44/42 MAPK was significantly activated by NE in non-epithelial cells (stromal cells and smooth muscle cells) while not in epithelial cells. The uptake of (3)H-thymidine was significantly increased by NE in both non-epithelial cells, which was inhibited by alpha1-adrenoceptor antagonists. CONCLUSIONS: These results suggest that NE may stimulate the proliferation of non-epithelial prostatic cells, which may be involved in the pathogenesis of BPH.

Clinicopathological study of renal cell carcinoma.

We investigated the clinicopathological features of 153 patients with renal cell carcinoma who underwent nephrectomy at our institute from April 1992 to August 2001 and evaluated the prognostic significance of various clinical and pathological factors. The prognostic factors were statistically analyzed by the log-rank and generalized Wilcoxon tests, and for the factors that were statistically significant by both analyses, multivariate analysis was conducted by stepwise regression in the Cox proportional hazard model. As a result, the overall 1-year, 3-year, 5-year and 10-year survival rates were 93.5%, 84.3%, 82.9% and 79.1%, respectively. In addition, multivariate analysis showed that regional lymph node metastasis, stage and histological type were independent prognostic factors for renal cell carcinoma. The number of renal cell carcinoma cases incidentally discovered during routine medical check-ups has increased in recent years, and in our study, such incidental tumors accounted for 65.3% of all cases. The 5-year survival rate of incidental tumors was 87.4% as compared to that of non-incidental tumors at 70.1%, and prognosis of incidental tumors was significantly more favorable. However, even in such cases, a favorable prognosis is not necessarily guaranteed according to the presence of regional lymph node metastasis, stage and histological type. Therefore, we conclude that even in incidental tumors, if the patient has unfavorable prognostic factors, post-operative adjuvant therapy should be considered.

Decreased sulfotransferase SULT1C2 gene expression in DPT-induced polycystic kidney.

BACKGROUND: The pathogenesis of polycystic kidney disease (PKD) remains unclear despite the identification of the genes responsible for hereditary PKD. In this study, we investigated the alteration of gene expressions in an acquired PKD model induced by 2-amino-4,5-diphenylthiazole (DPT) using the differential display method. METHODS: Kidney mRNA from a Sprague-Dawley rat fed with 1% DPT for 4 days and from a control rat was compared by the RT-PCR differential display method. Differentially expressed bands were re-amplified and subcloned. Using these subclones as probes, the changes in gene expressions were confirmed by Northern blot analysis. Subsequently, mouse kidney cDNA library was screened. RESULTS: The isolated 1.5-kb cDNA contained an open reading frame encoding 296 amino acids, which shared 94.3% identity with rat SULT1C2 sulfotransferase, and was considered to be its mouse ortholog (GenBank Accession No. AY005469). Mouse SULT1C2 mRNA was abundant in the kidney and stomach among normal mouse tissues. The expression of SULT1C2 mRNA was decreased in the rat kidney after DPT feeding but not in the stomach. Mouse SULT1C2 was expressed successfully using pET plasmid vector and E. coli. The recombinant 34-kD protein was capable of catalyzing the sulfation of p-nitrophenol at a Km of 3.1 mmol/L, by utilizing 3'-phosphoadenosine 5'-phosphosulfate (PAPS) as the sulfate donor. CONCLUSIONS: Although the physiological substrate and function of SULT1C2 have yet to be elucidated, its down-regulation could be involved in the cystic changes of tubules by decreasing the sulfation of the tubular basement membrane components.

Comparative antitumor activity of 5-fluorouracil and 5'-deoxy-5-fluorouridine in combination with interferon gamma in renal cell carcinoma cell lines.

The treatment of advanced renal cell carcinoma (RCC) remains unsatisfactory, and conventional therapy using interferon (IFN) has shown limited antitumor action. In this study, we examined the combined effects of IFN gamma with 5-fluorouracil (5-FU) as well as 5'-deoxy-5-fluorouridine (5'-DFUR), a prodrug of 5-FU, in vitro. The sensitivity of 4 RCC cell lines to IFN gamma and 5-FU or 5'-DFUR was evaluated using the MTT assay. The IC50 value for the cytostatics was expressed as the concentration at which growth was inhibited by 50% as compared with the control value. Thymidine phosphorylase (TP), the enzyme that converts 5'-DFUR into 5-FU and 5-FU into 5-fluoro-2'-deoxyuridine-5'-monophosphate (FdUMP), was estimated by ELISA. As a result, a significant correlation between TP expression and sensitivity to 5-FU and 5'-DFUR was found in all 4 RCC cell lines. By adding 12.5 U/ml of IFN gamma, which is a concentration that does not affect cell proliferation, the TP expression significantly increased by 1.5 to 3.4 times in the 4 RCC cell lines. Susceptibility to 5-FU and 5'-DFUR was also significantly induced by 1.42 to 2.36 times and 2.9 to 5.71 times, respectively, in the 4 cell lines. These results suggested that TP may be useful as a predictive factor in combination therapy with IFN gamma and 5-FU or 5'-DFUF, which may be a promising treatment for advanced RCC.