Combination therapy for renal cell carcinoma using IFN-gamma and OK-432: in vitro study.

Synergistic effect of recombinant IFN-gamma (IFN-gamma) and OK-432, a streptococcal preparation, using chromium release assay was studied in vitro on killer cell induction. The target cells utilized for assay were a human leukemia cell line K562, a human renal carcinoma cell line KU-2, autologous normal kidney tissues and autologous renal cell carcinomas. Culture supernatant of peripheral blood lymphocytes (PBL) and OK-432 (designated as OK conditioned medium or OK-CM) demonstrated enhanced cytotoxicity of fresh PBL against these target cells. Killer cell activity against autologous cancer cells could be also induced from PBL of renal cell carcinoma patients. Pretreatment of PBL with IFN-gamma revealed synergistic effect of OK-CM on killer cell induction. OK-CM derived from patients was shown to contain IL-2 activity as well as high titer of interferon. Neutralizing monoclonal antibody against IFN-gamma and IL-2 receptor demonstrated reduction of cytotoxicity. These results suggested potential benefit of sequential use of IFN-gamma and OK-432 for the treatment of metastatic renal cell carcinoma.

Human prostatic cancer cell derived growth factor stimulating fibroblasts and its mechanisms.

A human prostatic cell line JCA-1 has been shown to release into culture medium an activity that promotes the proliferation of murine fibroblast 3T3 cells. The presence of additional serum-free conditioned culture supernatant from JCA-1 cells accelerates the 3T3 growth rate as determined by cell counts, [3H]thymidine uptake into DNA and colony formation in soft agar gel. The growth activity was determined not to derive from culture medium components. Treatment of JCA-1 cells with RNA synthesis inhibitor actinomycin D abrogated the activity, indicating that protein synthesis is required for the presence of this activity from JCA-1 cells. The molecular mass of this prostatic cell derived growth factor (PRGF) was larger than 10,000 KDa, and its interaction with fibroblasts resulted in an approximate 27 percent increase in replicating cell cycle phases S and G2M with a reciprocal decrease of G1 cells. These data indicated a mechanism causing a more rapid entry of cells into replication. The interaction between prostatic cancer cells and related tissues and their relation to pathogenesis is reviewed.

Augmentation of cell-mediated cytotoxicity against renal carcinoma cells by recombinant interleukin 2.

The effects of recombinant interleukin 2 (IL-2) on natural killer (NK) cell activity against established renal carcinoma cell lines CaKi 1, KU-2, and freshly prepared renal carcinoma cells were studied. Augmentation of NK cell activity by IL-2 was dose- and time-dependent. The results indicate that the optimal dose of IL-2 was 100 to 500 U/mL, and that cytotoxicity increased significantly even at a low concentration such as 4 U/mL. IL-2 induced significantly higher levels of cytotoxicity against renal carcinoma cells than did gamma-interferon. The influence of IL-2 on lymphocyte subpopulations was then examined using flow cytometry with monoclonal antibodies OKT3, OKT4, OKT8, Leu 7, and Leu 11. The results showed that IL-2 increased the number of cells positive to Leu 11, the so-called active NK cells. We concluded that IL-2 has an important role in the treatment of renal carcinoma.

New Cavitron system (CUSA/CEM): its application for kidney surgery.

A new Cavitron Ultrasonic Surgical Aspirator system (CUSA/CEM), which contains electrocautery function at the hand piece, was used for partial nephrectomies in dogs without clamping the renal artery. The advantages of this new device (CUSA/CEM) over the original CUSA are: (1) it enabled more precise resection of the tissue in a dry field, (2) shorter operating time, and eventually (3) helpful for reduction of blood loss. The microscopic examination of the resected kidney revealed minimal reaction and damage to the adjacent kidney tissue as compared with the effect by electrocautery or microwave tissue coagulator.

Immunosuppressive factor derived from renal cancer cells.

An immunosuppressive factor derived from conditioned medium of a human renal cancer cell line, KU2, was analyzed, using mitogen-activated normal peripheral blood lymphocytes. Addition of conditioned medium to lymphocyte cultures resulted in suppression of [3H]-thymidine incorporation to DNA of lymphocytes in a dose-dependent manner. This effect was cytostatic and reversible. This factor inhibited lymphocyte activation stimulated by mitogen at an early stage, and it also suppressed Interleukin-2 production by activated lymphocytes. This factor was non-dialyzable and heat sensitive at 56 degrees C. DNA replication of the cells was necessary for the production of this factor.

Application of microwave tissue coagulation in partial nephrectomy.

Microwave tissue coagulation was used during partial nephrectomy in 10 mongrel dogs, without clamping the renal artery. There were no major complications, such as retroperitoneal hematoma, abscess formation, or macroscopic infarction of the kidney tissue related to this new procedure. The advantages of microwave coagulation are reduced blood loss, shorter operative time, and minimal risk of vascular injury.

Immunologic study of human recombinant interleukin-2 (low-dose) in patients with advanced renal cell carcinoma.

Immunologic and antitumor effects of human recombinant interleukin-2 was studied in patients with advanced renal cell carcinoma. Intravenous drip infusion for inpatients or subsequent subcutaneous injection for outpatients at a daily dose of 1 x 10(6) units of interleukin-2 was given over a period varying from twenty-one to two hundred forty days to 13 patients. Natural killer cell activity increased 20 percent or more in 11 of the 13 patients, and lymphokine-activated killer cell activity increased 40 percent or more in 9 of the patients tested. Interleukin-2-receptor positive cells and HLA-DR positive cells in peripheral blood lymphocytes increased in 12 and 11 patients, respectively. Two patients achieved complete response; 1 had a partial response; 6 had no change; and 2 had progressive disease. Two patients were not evaluated because the therapy had been discontinued in less than four weeks. We conclude that low-dose interleukin-2 enhances patients' immunity and has a potential antitumor activity against renal cell carcinoma.

Establishment of new human prostatic cancer cell line (JCA-1).

The establishment of a new human prostatic cancer cell line is described. This cell line was derived from a poorly to moderately differentiated prostatic adenocarcinoma. It has been maintained in tissue culture for fourteen months and has been passed fifty-two times. This cell line has an ability to form colonies in soft agar suspension cultures, and also is transplantable to nude mice. Tumors grown in nude mice revealed a poorly differentiated adenocarcinoma with positive PSA staining. Acid phosphatase activity was detected in freeze-thawed cells by enzymatic assay. A karyotype analysis demonstrated aneuploidy with a model chromosomal number of 69 and six marker chromosomes.

[A case of transitional cell carcinoma of renal pelvis with extremely high serum levels of CA19-9 and CEA].

We report a case of transitional cell carcinoma of the renal pelvis with extremely high serum levels of carbohydrate antigen 19-9 (CA19-9) and carcinoembryonic antigen (CEA). A 69-year-old woman was admitted with gross hematuria. Imaging diagnosis revealed a tumor in the left renal pelvis and multiple liver and bone metastases. Serum levels of CA19-9 and CEA were 2,557 U/ml (normal < 36) and 523 ng/ml (normal < 3.8), respectively. She died of cancer progression 3 weeks after admission. An autopsy diagnosis was transitional cell carcinoma of the left renal pelvis. No abnormal findings were recognized in the gastrointestinal organs. Cancer cells showed a positive immunohistochemical staining for both CA19-9 and CEA.

[Establishment of two renal cancer cell lines].

Two renal cell carcinoma (RCC) cell lines, JMSU2 and JMSU3, derived from the primary sites of mixed cell type and spindle cell type RCC, respectively, have been established and maintained for 31 and 22 months. Karyotypic analysis revealed human karyotypes with modal numbers of 84 and 55, respectively. Consistent chromosomal abnormalities were 1p+, 3p-, 6q- or 8p- in the JMSU2 cells and 1p-, inv (5p + q-) or loss of sex chromosome in the JMSU3 cells. Electron microscopy revealed abundant glycogen granules, lipid droplets and microvilli. The JMSU3 cells transplanted to nude mice produced tumors with a spindle cell pattern similar to that of the original tumor. High concentrations of cytokines, such as interleukin-6 (145,000 pg/ml), interleukin-8 (35,300 pg/ml) and granulocyte-colony stimulating factor (6,340 pg/ml), were detected in the culture supernatant of the JMSU3 cells. Interleukin-1 beta (IL-1 beta) dose-dependently inhibited the proliferation of the JMSU2 and JMSU3 cells in culture. Tumor cytotoxic factor/hepatocyte growth factor (TCF/HGF) dose-dependently enhanced JMSU3 cell proliferation, but suppressed JMSU2 cell proliferation. These findings suggest that IL-1 beta and TCF/HGF have regulatory roles in the proliferation of RCC.

[Percutaneous removal of renal and upper ureteral stones: clinical results and complications of 103 cases].

We report the results and complications of 103 consecutive patients who underwent percutaneous removal of renal and ureteral stones. The overall clinical success rate was 80.6%. For the recent 33 cases in which UL-arm fluoroscopy was used, however, the success rate was as high as 87.9%, which was considered to be due to easier establishment of percutaneous direct access. The most common complications were bleeding (18.5%), extravasation (15.5%) and fever (9.7%). Four cases with significant bleeding required arteriography, but there were no sign of arteriovenous fistula nor pseudoaneurysms in any cases. To study renal parenchymal damage in the percutaneous procedures, plasma renin activities (PRA) were compared in 54 cases after six months. However, significant elevation of PRA did not occur in any case.

[Lymphadenectomy in the operative treatment of renal pelvic and ureteral carcinoma].

We evaluated 18 patients with renal pelvic and ureteral carcinoma focusing on the operative treatment. All the patients underwent nephroureterectomy and partial cystectomy, except for two patients, with CIS of the ureter and renal insufficiency, and one with invasive bladder carcinoma, in whom partial ureterectomy and total cystectomy were performed, respectively. Lymphadenectomy were performed in all the patients and resected lymph nodes were from renal hilus, para-aorta or vena cava, according to the affected side, intra aorto-caval and all the pelvic nodes in the affected side. Lymph node metastases were found in 4 patients (22%), 2 in the primary and distant nodes and the other in the regional nodes only. Skipped lesion of the lymph node metastasis was not rare and lymph node dissection from the renal hilus to the pelvis on the affected side should be included in the operative treatment of renal and ureteral carcinoma.

[Demonstration of interleukin-1 (IL-1) as an autocrine growth inhibitor in renal cancer cell line, TC-1].

In this report, we demonstrated that Interleukin-6 (IL-6) production could be induced by stimulating renal cell carcinoma cell lines, namely ACHN, Caki-1 and TC-1 cells with Interleukin-1 beta (IL-1 beta). IL-1 beta had no effects on cell proliferation in ACHN cells. However, IL-1 beta could suppress cell proliferation in Caki-1 and TC-1 cells. Flow cytometric cell cycle analysis by double staining method with propidium iodide and Proliferating cell nuclear antigen (PCNA) disclosed IL-1 beta caused cell accumulation at G1 phase. Fine granules were visualized in perinuclear area of TC-1 cells treated with IL-1 beta under microscopy. High electron density granules and spherically dilated rough endoplasmic reticula were observed by electron microscopic examinations. In TC-1 cell culture, IL-1 beta excretion into the supernatant was demonstrated by bioassay and ELISA. These results suggest that IL-1 beta functions as an "autocrine growth inhibitor" against TC-1 cells. Half-maximal inhibition of IL-1 beta and IFN-alpha was 6.5 pg/ml, and 720 U/ml, respectively for TC-1 proliferation and combination of these cytokines showed enhanced activity in cell growth inhibition.

[A case of teratoma of the testis with retroperitoneal lymph nodes involved with malignant teratoma after chemotherapy].

This report describes a case of teratoma with retroperitoneal lymph nodes involved with malignant teratoma (enteric adenocarcinoma) after extensive chemotherapy for the original testicular cancer. A 18-year-old man with a mixed cell tumor (embryonal carcinoma+teratoma+yolk sac tumor) received three courses of VAB-6 chemotherapy for bulky mass following inguinal orchiectomy. He was referred to Tochigi Cancer Center for treatment of a residual mass. He was treated with resection of the mass combined with left nephrectomy due to severe adhesion and pathological diagnosis of the resected lymph node was mature teratoma with a massive necrotic tissue. Two courses of BEP chemotherapy were given to the patient following the surgery. Six months after completion of chemotherapy, a retroperitoneal mass of 1.5 cm in diameter, was detected by CT scan. Standard retroperitoneal lymph node dissection was performed and the pathological diagnosis of the lymph node was teratoma with malignant transformation containing enteric adenocarcinoma. Teratomatous portion of the primary lesion was precisely re-examined and adenocarcinoma, similar histology to the retroperitoneal mass, was identified. He received two courses of EAP chemotherapy (Cis-platin+etoposide+doxorubicin) as an adjuvant chemotherapy following the surgery and he is alive with no evidence of recurrence for 21 months. Presence of non germ cell malignancy after chemotherapy in testicular cancer has been regarded as a rare phenomenon. Flow cytometric DNA analyses of both embryonal carcinoma and teratoma in the primary lesion, mature teratoma and teratoma with malignant transformation of the retroperitoneal lymph node disclosed that these tumors were all aneuploid tumors.(ABSTRACT TRUNCATED AT 250 WORDS)

[Establishment of a new human renal cancer cell line (TC-1) and its productivity of interleukin-6 (IL-6)].

We established a new cell line (TC-1) from primary site of a renal cell carcinoma (RCC) patient. Its doubling time in tissue culture was 20 hours at 45th passage and mycoplasma contamination test was negative. The karyotypic analysis demonstrated a human karyotype with a modal number of 70. A consistent chromosomal abnormality was noted such as No. 4 monosomy, No. 7 trisomy and a loss of Y chromosome. Electron microscopic examination showed a brush border, vacuoles and abundant glycogen granules in the cytoplasm, which was compatible with RCC cells. This cell line was transplantable to nude mice and the grown tumor closely resembled the original tumor, i.e. clear cell type and hypervascularity. High titer of interleukin-6 (IL-6) was detected in the supernatant of TC-1 cell culture (approximately 5 ng/ml) as well as in sera of nude mice bearing this tumor (260 pg/ml). Exogenous IL-6 did not enhance the TC-1 cell proliferation as determined by cell count. Flow cytometric analysis could not demonstrate the existence of IL-6 receptor on the cell surface. These results suggested the produced IL-6 did not act as an autocrine growth factor in the cell line. Additional IL-1 alpha to the culture medium induced 3-4 times higher concentration of IL-6 in the culture supernatant compared with that of non-stimulating cells, while exogenous TNF alpha did not stimulate IL-6 production.

[Production of a lymphocyte chemotactic factor by human renal cancer cells].

Tumor infiltrating lymphocyte (TIL) is commonly observed in renal cell carcinoma (RCC) tissues. First step of lymphocyte accumulation in cancerous tissues is the lymphocyte migration toward cancer cells. However, no conclusion has been drawn which cytokine is involved in TIL of RCC. The purpose of this study is the identification of a lymphocyte chemotactic factor (chemokine) produced by a newly established RCC cell line. A new human renal cancer cell line (TC-2) was established from a primary site of a 52-year-old man. A marked lymphocyte infiltration was noticed at the cancer tissue. A tissue culture has been continued for 24 months. Flow cytometric analysis of this cell line revealed DNA aneuploidy. A human karyotype, with a modal number of 72, and consistent abnormalities, such as 4q+ and 5q-, were demonstrated by Giemsa banding analysis. Approximately 2.9 fold of lymphocyte chemotactic activity was detected in the culture supernatant of TC-2 cells (TC-2CM) as measured by in vitro migration assay. Sixty percent of this activity was abrogated by adding the neutralizing antibody against interleukin-8 (IL-8) into TC-2CM. Analysis of surface markers of migrating lymphocytes disclosed that lymphocytes expressing CD3, CD8 and CD16 phenotype predominantly showed migration. These results suggested that chemotactic activity for lymphocytes derived from TC-2 cells was partly IL-8.

[Effects of recombinant interleukin 1 beta on peripheral blood cells and induction of lymphokine-activated killer activity in patients with urological malignancies].

Subcutaneous injection of human recombinant interleukin 1 (IL-1) beta was given to 9 patients with urological malignancies (5 renal cell carcinoma, 2 bladder carcinoma, 1 renal pelvic tumor, and 1 testicular tumor), at an initial dose of 1 x 10(4) units on days 1 and 2, and there after weekly for 4 weeks. The dose was increased by 1 x 10(4) units weekly up to final dose of 4 x 10(4) units. Peripheral blood mononuclear cells (PBMC) were isolated from patients on day 3 in week 2 and week 4, and lymphokine-activated killer (LAK) activity against Daudi cells was measured using 4 hr 51Cr-release assay, after incubation with human recombinant interleukin 2 (IL-2) of 50 units/ml for 72 hours. Proliferation of lymphocytes was measured by tritiated thymidine incorporation after incubation with IL-2 for 72 hours. IL-1 beta increased the number of peripheral blood granulocytes and lymphocytes, but did not increase the numbers of monocytes and platelets. IL-1 beta significantly augmented IL-2-induced LAK activity in vitro, but this augmentation was neither accompanied by the increase of IL-2 receptor-positive cell ratio in peripheral blood lymphocytes nor enhancement of IL-2-induced proliferation of lymphocytes. Administration of IL-1 beta increased LAK activity of the patients, despite the fact that IL-1 beta did not increase LAK activity in vitro. The result suggests that IL-1 beta-stimulated LAK activity may be mediated by the induction of some cytokines in the patients.(ABSTRACT TRUNCATED AT 250 WORDS)

[A case of porocarcinoma arising on the penile shaft].

BACKGROUND: Porocarcinoma is a rare eccrine sweat gland tumor, and a varied treatment is not yet determined for metastatic disease because of the chemo-resistant and radio-resistant nature of this tumor. CASE: This report describes a case of porocarcinoma arising on the penile shaft of 83-year-old man with extensive lymph node metastases. He was treated with emasculation and bilateral ilio-inguinal lymph node dissection. Histological examination of the resected tumor showed a homogeneous round cells with duct-like formation in some part and Pagetoid infiltration was also noted in the dermis. Immunohistochemical examination demonstrated a positive CEA and negative S-100 staining and this result was consistent with the pathological diagnosis of porocarcinoma. One course of chemotherapy which consisted of methotrexate, cisplatin, adriamycin, and bleomycin was given to the patient following surgery for the treatment of residual lymph nodes in the paraaortic area. Abdominal CT scan revealed partial response, about -94% shrinkage, after one course of chemotherapy. The duration of the response lasted four 4 months. He died of pneumonia caused by MRSA after 4 months and the autopsy revealed multiple liver metastases and a massive infiltration of tumor cells in the bone marrow of lumbar vertebra. CONCLUSIONS: There is no definite treatment modality available for metastatic porocarcinoma. Because of the patient's age, only one course of chemotherapy was given, however, a fairly good response against this rare tumor suggested that this new regimen might be effective against porocarcinoma.

[Role of serum E-selectin (ELAM-1) and inflammatory parameters in patients with renal cell carcinoma].

(BACKGROUND). E-selectin is an adhesion molecule expressed on IL-1 activated endothelial cells and it binds to carbohydrate ligands such as sialy Lewis A antigen (SLeA) or Lewis X antigen (SLeX) on cancer cells. This mechanism is supposed to play an important role during hematogenous metastasis. Some of renal cell carcinomas (RCC) are known to produce inflammatory cytokines such as IL-1 beta and IL-6 and clinical evidence shows that the prognosis of this type of tumor is generally poor. We investigated whether this adhesion molecule was involved in hematogenous metastasis. (METHOD). In the present study, soluble E-selectin level was measured in the sera of 89 patients with RCC prior to nephrectomy or IFN treatment using sanwich ELISA method. (RESULTS). The results indicated that high E-selectin concentration in the patients' sera was correlated with low incidence of metastasis and consequently correlated with good prognosis of RCC patients. Inflammatory serum parameters, such as serum C reactive protein (CRP), immunosuppressive acid protein (IAP) and erythrocyte sediment rate (ESR) were also assessed and these parameters were revealed to be negatively correlated with the serum level of E-selectin. In order to investigate this mechanism, we performed in vitro study on RCC cell/endothelial cell adhesion. IL-1 beta enhanced adhesion of 2 RCC cell lines and this adhesion was partially inhibited by adding exogenous E-selectin into the culture medium. Expression of SLeA and SLeX were demonstrated on the cell surface of 2 RCC cell lines by flowcytometric analysis. (CONCLUSION). The results suggested that E-selectin and SLeX/SLeA interaction was involved in the adhesion between RCC and endothelial cells and also inflammatory cytokine production by RCC cells was a risk factor for metastasis through E-selectin induction. Although expression of E-selectin on endothelial cells facilitates metastasis, excessive production of E-selectin into the serum was suggested to have inhibitory effect against metastasis.

[Primary adenocarcinoma of the tunica vaginalis testis expressing CA19-9 antigen].

A 47-year-old man presented with a 10-year history of left scrotal swelling. No solid mass detected on sonogram, dark-red colored fluid aspirated, equivocal cytology, and elevated serum CA19-9 level prompted surgical exploration. Frozen section reported no malignancy of thickening of the tunica vaginalis. The final pathology adenocarcinoma, however, necessitated left radical orchiectomy and hemiscrotal resection, demonstrating no malignancy at the left testis nor epididymis. CA19-9 level was high (104,200 U/ml) in the fluid obtained at the first surgery and the tumor cells were positively stained for CA19-9. Imaging study including abdominal CT scan and upper G-I series and normalization of CA19-9 level denied other tumor existing. In conclusion, this is to our knowledge the first case report of adenocarcinoma of the tunica vaginalis expressing CA19-9 antigen.