Antitumor effect of reduction of 150-kDa oxygen-regulated protein expression in human prostate cancer cells.

Heat shock proteins (HSPs)/stress proteins are molecular chaperones that are induced by various environmental and physiological stimuli. Evidence of the relations between the expression of HSPs and the regulation of cell growth or transformation has accumulated. The 150-kDa oxygen-regulated protein (ORP150), a new member of HSP family, functions as a molecular chaperone in the endoplasmic reticulum. We have examined whether transduced antisense ORP150 cDNA reduces tumorigenicity and angiogenicity. Relations between these stress proteins and cancer and possibilities for anticancer gene therapy are described.

[Primary small cell carcinoma of the kidney: a case report].

A 43-year-old male visited our hospital with the complaint of right flank colicky pain. Computed tomographic (CT)-scan and angiography showed large renal tumor with liver invasion and tumor thrombosis in the vena cava. Multiple lung and bone tumors were also recognized. Percutaneous biopsy of the renal tumor revealed small cell carcinoma. Multiple lung masses were diagnosed as metastatic tumors according to the results of bronchoscopic biopsy. Chemotherapy including cisplatinum and etoposide was performed without success. He died 6 months after the diagnosis. Autopsy specimen revealed primary small cell carcinoma of the right kidney. To our knowledge, this is the seventh case as primary renal small cell carcinoma in the world literature.

[A case of adrenal pheochromocytoma with contralateral adrenocortical adenoma].

A case of a pheochromocytoma in the right adrenal gland and adrenocortical adenoma in the left adrenal gland of a 58-year-old male is reported. The patient was incidentally found to have a right adrenal tumor by ultrasonographic study. A computerized tomographic (CT) study and magnetic resonance image (MRI) study revealed bilateral adrenal tumors. The sizes of the right tumor and left tumor were 2.5 x 3.5 cm and 1.2 x 1.0 cm, respectively. The intensity of each tumor was different on T2-weighted MRI. 131I-MIBG scintigram showed the uptake of right adrenal gland. The existence of pheochromocytoma was confirmed by the elevated levels of catecholamines. We performed venous sampling to be certain whether the patient had unilateral or bilateral pheochromocytoma. As a result, bilateral adrenal pheochromocytoma was diagnosed. Therefore, we performed bilateral adrenalectomy. However, histopathological examination revealed right pheochromocytoma and left non-functioning adrenocortical adenoma.

Juxtaglomerular cell tumor of the kidney treated with nephron-sparing surgery.

A case of juxtaglomerular cell tumor of the right kidney is reported. A 30-year-old woman visited us with a complaint of headaches. Severe hypertension and an elevated level of plasma renin activity was seen at the initial evaluation. Computerized tomographic angiography revealed tumor vessels in the low-density mass in the right kidney. The preoperative diagnosis was renin-secreting tumor of the kidney, and nephron-sparing surgery was performed. The pathological findings showed a juxtaglomerular cell tumor. Postoperatively, prompt normalization of blood pressure and a reduced plasma renin activity level were observed.

[A case of chromophobe cell renal carcinoma].

We report a case of chromophobe cell renal carcinoma. A 27-year-old male suffered from right flank pain and gross hematuria. Drip infusion pyelography and ultrasonographic examination revealed a right renal mass and it was diagnosed as renal oncocytoma by needle biopsy of the right kidney. He was referred to our hospital for the operation and right radical nephrectomy was performed. The cut surface of the tumor was beige in color. The cytoplasm of the tumor cells stained positively for colloidal iron and showed a negative reaction for Vimentin. From these results, this tumor was given a diagnosis of chromophobe cell renal carcinoma.

[Correlation between expression of metastasis-related genes and lymph node metastasis in testicular cancer].

To investigate the factors related to lymph node metastasis of testicular germ cell tumors, we first established a seminoma orthotopic model with lymph node metastasis in SCID mice by inoculating small fragments from subcutaneous xenografts. Second, we compared the expression patterns of metastasis-related genes of the seminoma xenografts and of the TCam-2 cells which were established as a seminoma cell line from a primary testicular seminoma. Third, we immunohistochemically analyzed human germ cell tumors (25 seminomas, 17 nonseminomas) using monoclonal antibodies to CD34, VEGF, VEGF-C, Flt-4, MMP-2 and E-cadherin. Testicular seminoma xenografts grew in 32/32 (100%) of the inoculated mice, of which 15 (47%) developed macroscopic metastasis to the renal hilar lymph node. Circulating tumor cells were detectable by using a PCR assay for the human beta-globin gene in 25/32 (78%) mice, although metastatic foci were not histologically evident in the visceral organs, including lungs, liver, kidneys and spleen. This may reflect the lymphophilic characteristics of the seminoma cells used. Regarding mRNA expression of metastasis-related genes, an increased expression of MMP-2 and VEGF compared with that in the s.c. xenografts was demonstrated by RT-PCR assay in the testicular seminoma xenografts. In addition, uPAR, MMP-1, MMP-2, MT1-MMP and MT3-MMP showed a a stronger expression and PAI-2 a weaker expression in the seminoma xenografts than did TCam-2 cells. These results suggest a higher metastatic potential of the seminoma xenografts, especially testicular xenografts, as compared with TCam-2 cells. In the immunohistochemical study, a significant correlation was found between MMP-2 expression and lymph node metastasis, which is compatible with the results for the metastasis-related gene expression from the seminoma xenografts.

[Renal cell carcinoma recurrence in the mediastinum lymph node 19 years after nephrectomy: a case report].

A 62-year-old male, who had undergone right nephrectomy to treat renal cancer 19 years earlier, was recently referred to our hospital to receive a detailed examination and treatment of mediastinal lymph node swelling. Biopsy of the swollen lymph nodes allowed a diagnosis of renal cell carcinoma (alveolar type, clear cell subtype, GI) to be made. The pathological features of his tumor were consistent with those of the renal tumor resected 19 years previously. Because there was a high probability of further growth of the swollen mediastinal lymph nodes and consequent high probability of compression of the superior vena cava, we performed mediastinal lymph node excision. Immediately after surgery, prophylactic interferon therapy was started. To date, five cases (including the present case) in which renal tumors recurred more than 15 years after surgical treatment have been reported in Japan.

Expression of messenger RNAs for membrane-type 1, 2, and 3 matrix metalloproteinases in human renal cell carcinomas.

PURPOSE: Three different membrane-type matrix metalloproteinases (MT1, 2, and 3-MMP) that can activate proMMP-2 (progelatinase A) are thought to play an important role in invasion and metastasis by various human carcinomas. To further clarify this role, we examined mRNA expression of MT-MMPs in human renal cell carcinomas. MATERIALS AND METHODS: mRNA was extracted from 25 clinical specimens of renal cell carcinoma and 23 specimens of normal renal parenchyma remote from the tumor. Reverse transcription polymerase chain reaction (RT-PCR) using specific primers was performed, and PCR products were hybridized to 32P-labeled internal probes and analyzed by a bioimage analyzer. RESULTS: MT1, 2, and 3-MMP mRNA expression in carcinomas was significantly higher than in normal parenchyma. In terms of the pathologic stage, MT1-MMP mRNA expression in pT2 and pT3 tumors was significantly higher than those in pT1 tumors. Although the sample size was small, it was evident that MT3-MMP mRNA expression in clear cell subtype renal cell carcinomas was higher than in the group of tumors including the granular cell subtype. CONCLUSIONS: These three MT-MMPs may play an important role in the pathogenesis of human renal cell carcinoma, and MT1-MMP in particular is important in invasion by carcinoma cells. It is interesting that the expression of MT3-MMP was higher in carcinomas, especially clear cell carcinoma, than in normal parenchyma, so that MT3-MMP may provide a clue an understanding of the molecular mechanism of carcinogenesis in human kidney.

A human seminoma xenograft model with regional lymph node metastasis.

PURPOSE: To establish a seminoma orthotopic model with lymph node metastasis to investigate the factors related to the lymphophilic behavior of seminoma cells. MATERIALS AND METHODS: Testicular seminoma xenografts were established by the inoculation of small fragments from subcutaneous (s.c.) xenografts that had previously been established in severe combined immunodeficient (SCID) mice with a supraclavicular lymph node metastasis from a human seminoma. Hematologic dissemination of tumor cells was analyzed by polymerase chain reaction (PCR) amplification of the human beta-globin gene. Xenograft messenger RNA levels of metastasis-related genes were examined by reverse transcription (RT)-PCR. RESULTS: Testicular seminoma xenografts grew in 32/32 (100%) of the inoculated mice, of which 15 mice (47%) developed macroscopic metastasis to the renal hilar lymph node. Circulating tumor cells and tumor cell shedding in the lung and liver were detectable by PCR assay in 25/32 (78%), 32/32 (100%), and 27/32 (84%) mice, respectively, although metastatic foci were not histologically evident in these organs. Increased expression of matrix metalloproteinase-2 (MMP-2), membrane-type 3 matrix metalloproteinase (MT3-MMP) and vascular endothelial growth factor (VEGF), and reduction in expression of plasminogen activator inhibitor-2 (PAI-2) were demonstrated by RT-PCR assay in the testicular xenografts as compared with the s.c. xenografts. CONCLUSIONS: This model mimics the lymphophilic behavior of seminoma and may help in elucidating the molecular mechanism of tumor spread via the lymphatics.

Dominant role of E-cadherin in the progression of bladder cancer.

PURPOSE: To elucidate factors with a role in the progression of bladder cancer. MATERIALS AND METHODS: One invasive (T24) and two superficial (RT4 and KK47) human bladder cancer cell lines, which express metastasis-related genes, were used. Cells were intravenously inoculated into chick embryos to evaluate metastatic potential to the liver. An orthotopic model with severe combined immunodeficiency mice was also used to investigate both histological appearance and changes in metastasis-related gene expression. Finally, gene expression patterns in a clinical setting were compared between superficial and invasive bladder cancers. RESULTS: In culture condition metastasis-related genes, including matrix metalloproteinases, urokinase-type plasminogen activator, and integrins alpha2 and alpha3 were continually expressed in T24 but only slightly or not at all in RT4 and KK47, respectively. The expression pattern of the metastasis-related genes in vitro reflected the characteristics of the original tumors. Liver metastasis in chick embryos was demonstrated not only with T24 cells, but also with RT4 cells in which enhanced expression of metastasis-related genes was induced. In the orthotopic model, histological appearances were in accordance with the characteristics of the original tumors, although enhanced gene expression was notable with RT4. Expression of E-cadherin by Western blotting was demonstrated only with RT4 under these experimental conditions. Furthermore, predominant E-cadherin mRNA expression was found in superficial and not in invasive human primary bladder cancers; expression of other genes was similar in the two groups. Dominant expression of E-cadherin in superficial tumors was confirmed by immunohistochemical analysis. CONCLUSIONS: These results implicate loss of E-cadherin expression as a critical factor in facilitating the progression of bladder cancer.

Expression and tissue localization of membrane-types 1, 2, and 3 matrix metalloproteinases in human urothelial carcinomas.

PURPOSE: Three different membrane-type matrix metalloproteinases (MT1, 2, 3-MMP) which can activate proMMP-2 (progelatinase A) are thought to have an important role in various human carcinoma invasions and metastases. We examined the mRNA expression of MT-MMPs and the tissue immunolocalization of MT1-MMP in human urothelial carcinomas. MATERIALS AND METHODS: mRNA was extracted from 27 clinical urothelial carcinomas and 10 normal urothelial mucosa tissues remote from the tumor. RT-PCR using specific primers was performed, and PCR products were hybridized to 32P-labeled internal probes and analyzed by a bioimage analyzer. Immunolocalization was studied using a monoclonal antibody against MT1-MMP (114-6G6). RESULTS: MT1-MMP and MT2-MMP mRNA expressions in urothelial carcinomas were significantly higher than those in the normal mucosa. In contrast, MT3-MMP mRNA was little expressed in both tissues, and the amount of MT3-MMP mRNA appeared to be much lower than MT1-MMP and MT2-MMP in the tissue samples. In terms of the tumor multiplicity, MT1-MMP and MT2-MMP mRNA expressions in the group of multiple tumors were significantly higher than those in the solitary tumor group. The carcinoma cells were immunostained for MT1-MMP predominantly in invasive and superficial carcinoma cells. The immunoreactivity was more intense in the invasive type than in the superficial type. CONCLUSIONS: It is suggested that MT1-MMP and MT2-MMP play an important role in the development of human urothelial carcinomas and reflect some aspects of the pathogenesis of multifocal occurrence. In spite of the possible contribution to the invasive and metastatic phenotype, MT1-MMP mRNA and its product are thought to be expressed already in the clinical superficial stage in some cases of this tumor type.

A chick embryo model for metastatic human prostate cancer.

OBJECTIVE: To establish a suitable experimental model of bone and liver micrometastases from human prostate cancer for evaluating antitumor agents. METHODS: PC-3 cells, an androgen-independent prostate cancer cell line, were inoculated into the chorioallantoic membrane vein of 10-day-old chick embryos (10(6) cells/egg). The polymerase chain reaction product for the human beta-globin gene in chick embryo femur and liver was quantified at various time points after inoculation, when immunohistochemical staining was done for Ki-67 antigen and cytokeratin. The antitumor effect of suramin was evaluated using the model, as was regional blood flow after thallium injection. RESULTS: Micrometastases were identified in bone and liver 1 day after inoculation and grew to form established metastatic foci in all embryos. Suramin showed significant antitumor effect for liver metastases, but not for those in bone where blood flow was relatively low. CONCLUSION: The chick embryo system provides a highly reproducible model for bone and liver micrometastases from human prostate cancer, suitable for evaluating antineoplastic agents at an early stage of the metastatic process.

Induction of necrosis by zinc in prostate carcinoma cells and identification of proteins increased in association with this induction.

Zinc exhibits inhibitory effects on apoptosis, and a deficiency in this metal generally causes this type of cell death to occur. In the present study, we found that exposure to zinc results in necrosis of prostate carcinoma cells. When zinc acetate was added to LNCaP or PC-3 cells in monolayer culture, they began to detach from the culture dishes, and viability was lost after 4-8 h. Most of the cell death was found to be due to necrosis as determined by double staining with fluorescein-isothiocyanate-labeled annexin V and ethidium bromide, and by detection of hypodiploid cells. Associated with the induction of necrosis was an increase in low molecular-mass proteins, identified by HPLC analysis to be thymosin beta10, parathymosin and GAGE in LNCaP cells, and thymosin beta4, parathymosin and metallothionein in PC-3. The time course of the increase of thymosin beta10 in LNCaP cells and thymosin beta4 in PC-3 cells was consistent with that of appearance of cell detachment and dead cells. These results indicate that zinc can induce necrosis and suggest that production of proteins including beta-thymosins is involved in induction of processes leading to cell detachment.

[Pulmonary metastasis from renal cell carcinoma diagnosed by using thoracoscopic biopsy: a case report].

A 70-year-old woman underwent radical nephrectomy for right renal cell carcinoma and received prophylactic interferon-alpha (rHuIFN-alpha) administration for one year. Followup computerized tomography scan showed a small nodule in the right lung 39 months postoperatively. Pulmonary nodules had become multiple and increased in size (6 mm) at 53 months. To examine the pulmonary lesions, histopathologically thoracoscopic biopsy of the right pulmonary nodule was performed and the diagnosis of metastatic renal cell carcinoma was confirmed. Because of its minimal invasiveness, thoracoscopic biopsy may be indicated in selected cases.

Immunolocalization of anti-placental alkaline phosphatase monoclonal antibody in mice with testicular tumors and lymph node metastasis.

To evaluate the ability of an anti-placental alkaline phosphatase (PLAP) monoclonal antibody (MAb) to localize to PLAP-expressing tumors, we established a model of testicular tumor with metastasis to lymph nodes and liver in severe combined immunodeficient (SCID) mice. 131I-labeled or 125I-labeled MAb was simultaneously administered via the intravenous or lymphatic route, respectively. Preferential accumulation of MAb in PLAP-expressing tumors at primary as well as metastatic sites was demonstrated. The percentage of the injected dose of MAb found in the tumor was generally higher when MAb was administered intravenously. Identical tumor/blood ratios were found with the two routes of administration. These data suggest that intravenous administration of a radiolabeled MAb is superior to lymphatic administration for tumor imaging and radioimmunotherapy.

Prognostic significance of c-erbB-2 gene expression in the poorly differentiated type of adenocarcinoma of the stomach.

Prognostic significance of c-erbB-2 gene abnormalities is unclear in the poorly differentiated type of gastric carcinoma, because the abnormalities of this gene have been reported to be restricted to the differentiated type of gastric carcinoma. In this study, correlation of c-erbB-2 gene amplification/overexpression of mRNA and protein were studied in the poorly differentiated type of gastric carcinoma. c-erbB-2 gene amplification determined by the slot-blot hybridization was observed in 11 (13%) of 82 gastric cancer, and 8 of 11 tumors were poorly differentiated. In addition, c-erbB-2 mRNA expression was studied by the reverse transcriptase-polymerase chain reaction. Four (17%) of 24 tumors showed overexpression of c-erbB-2 mRNA, and all these four exhibited morphologically a poorly differentiated type. Among 157 poorly differentiated gastric cancers, 20 (13%) tumors showed immunohistochemically c-erbB-2 protein expression. These tumors had significantly higher incidences of larger tumor, serosal invasion-positive tumors, node-positive tumor, or peritoneal dissemination-positive tumor than those without c-erbB-2 expression. Furthermore, patients with c-erbB-2 protein overexpression ran poorer prognoses than those without c-erbB-2 expression. From these results, we conclude that expression c-erbB-2 tissue status may be a good prognostic indicator in poorly differentiated gastric carcinoma.

Chemotherapy of metastatic testicular germ cell tumors: relationship of histologic response to size reduction and changes in tumor markers.

BACKGROUND: Although testicular germ cell tumor is one of the most curable cancers, approximately 20% of advanced cases remain incurable. In this study we investigate factors that may predict a poor response to standard chemotherapeutic regimens and thus allow earlier initiation of more aggressive measures. METHODS: We analyzed the records of 19 patients with metastatic testicular germ cell tumors (8 seminomas and 11 nonseminomas). Sixteen patients underwent surgical exploration for residual tumors following chemotherapy, and the histological findings on the resulting specimens were correlated with reductions in tumor size observed on computed tomography and with changes in tumor marker levels. RESULTS: Complete necrosis was obtained in 10 of 12 lesions that shrank by at least 80%, while continued existence of teratoma or cancer was confirmed in 9 of 11 lesions with smaller size reductions. An initial human chorionic gonadotropin-beta subunit (HCG-beta) level more than 100 times the upper limit of normal appeared to predict poor histological response (teratoma/cancer) to chemotherapy. Slow fall (prolonged half-life) of tumor markers during chemotherapy also correlated with poor histological response. CONCLUSION: Factors which predict poor histological response of tumors to chemotherapy include size reduction less than 80%, initial HCG-beta levels more than 100 times the upper limit of normal, and prolonged half-life of tumor markers (placental alkaline phosphatase, alpha-fetoprotein and HCG-beta).

Natural interferon enhances expression of placental alkaline phosphatase in human seminoma xenograft.

The purpose of this study was to investigate the effect of natural interferon (IFN) on the expression of placental alkaline phosphatase (PLAP) in a human seminoma xenograft in severe combined immunodeficient mice. Mice were injected intramuscularly with 3x10(5) U/mouse of IFN, twice a day, for five consecutive days. A significant increase in PLAP level of the xenografts followed IFN treatment. A radiolabeled anti-PLAP monoclonal antibody (MAb) was intravenously injected on the first day of IFN administration in order to determine if IFN has the potential to enhance the efficacy of an anti-PLAP MAb in the detection of seminoma. Enhanced retention of the anti-PLAP MAb was observed at 7 and 11 days after MAb administration. Thus, IFN treatment appears to have some effect on the efficacy of the anti-PLAP MAb in the detection of seminoma xenografts.

[An experimental study of laser treatment using two different type of neodymium:YAG laser on transplanted tumor in Scid mice].

BACKGROUND: We report basic animal experimental study which was evaluated the thermocoagulation effects of two different type of Nd:YAG laser (pulse and continuous wave (CW) laser). METHODS: The Rotalase internally reflecting fiber delivery system coupled to the pulse and CW Nd:YAG laser was used to create lesions in Seminoma tissue implanted Scid mice with the fiber tip 2 mm away from the target tissue under the water. Laser power output used was 20, 40 and 60-watt for varying times (30-180 sec) of irradiation. Stationary lesions, where a single spot of target was irradiated, were created. On the other hand, we measured tissue temperatures at 7, 10 and 14 mm from tissue surface. The seminoma tissues removed from Scid mice were photographed after bisection, the fixed in 10% formalin and examined histologically. RESULTS: For the purpose of these experiments, ablated tissue is defined as the volume of tissue that has been destroyed by both coagulation and vaporization. Estimates of the volume of ablated tissue were made by macroscopic examination of the bisected lesion, measuring the depth and width of the lesion as seen from the edges of the coagulated area around vaporized zone. The mean depth and width penetration, volume ablation and rising of the tissue temperature at pulsed 60-watt, 60 seconds was greater than that observed at other groups. Irradiated spot lesions were characterized by an initial 10-20 second period of tissue blanching followed by an audible "popcorn phenomenon" which meant more than 100 degrees C in tissue temperature, then formation of small surface bubbles as tissue began to evaporate and char. CONCLUSION: This study suggests the potential usefulness of the pulse Nd:YAG laser for Visual Laser Ablation of the Prostate (VLAP).