Satoshi Fukata

Expression of Angiogenesis-Related Genes Regulates Different Steps in the Process of Tumor Growth and Metastasis in Human Urothelial Cell Carcinoma of the Urinary Bladder

Objective: This study was designed to determine the relative activity of angiogenesis-related genes in the regulation of tumorigenicity and subsequent metastases of urothelial cell carcinomas (UC) of the urinary bladder. Methods: We selected the clones with the highest and lowest expression level of basic fibroblast growth factor (bFGF), vascular endothelial growth factor (VEGF)/vascular permeability factor or interleukin-8 (IL-8) in the highly tumorigenic and metastatic human UC cell line 253J B-V. Tumorigenicity and production of spontaneous lymph node metastases were evaluated 1, 2, 4, 8 and 12 weeks after orthotopic implantation of each specific expression clone into the urinary bladder of athymic nude mice. Moreover, the transitional changes in the expression of angiogenesis-related genes and neovascularization were determined in tumors and metastases. Results: At the early stage of tumor growth following orthotopic implantation, tumorigenicity and metastases were significantly increased in the clones with the highest expression of bFGF and IL-8, while they were significantly inhibited in the clones with the lowest expression of bFGF and IL-8 compared to parental 253J B-V. In the tumors, specific expression of angiogenesis-related genes and intratumor neovascularity of each clone were gradually regulated to the same level as parental 253J B-V. In metastasized tumors of the highest and lowest IL-8-expressing clones, IL-8 expression was consistently high and low, respectively. Conclusions: These findings indicate that at the early stage of tumor growth, bFGF and IL-8 expression play important roles in the regulation of angiogenesis, tumorigenicity and subsequent metastases of human bladder cancer.

Effect of Combination Therapy with a Novel Bisphosphonate, Minodronate (YM529), and Docetaxel on a Model of Bone Metastasis by Human Transitional Cell Carcinoma

Purpose: Transitional cell carcinoma (TCC) of the urinary tract is a chemosensitive tumor. Most deaths from TCC of the urinary tract are caused by metastasis, which is resistant to conventional chemotherapy. Frequent sites of metastases from TCC of the urinary tract are regional lymph nodes, liver, lung, and bone. Of these distant metastases, bone metastasis is consistently resistant to cisplatin-based conventional chemotherapy. Therefore, in this study, we investigated whether or not a newly developed minodronate, YM529, could prevent osteolytic bone metastasis of human TCC and also enhance the effect of docetaxel in a bone tumor model of athymic nude mice. Experimental Design: In the present study, we evaluated the effect of in vitro treatment with minodronate and/or docetaxel on the proliferation by cell count, the induction of apoptosis by terminal deoxynucleotidyl transferase–mediated nick end labeling (TUNEL) assay, and the biological activity of osteoclast by pit formation assay in human bladder cancer cell line, UMUC-14, and mouse osteoclast cells. In vivo, we examined the effect of minodronate in a bone tumor model of athymic nude mice, in which the percutaneous intraosseal injection in the tibia of UMUC-14, leads to osteolytic bone tumor, as a bone metastasis model. To examine whether or not minodronate could inhibit tumorigenicity and enhance the effect of the chemotherapeutic agent, docetaxel, we gave minodronate i.p. and/or docetaxel i.p. to nude mice 3 days after an intraosseal tumor implantation. Moreover, proliferation and the induction of apoptosis of cancer cells and osteoclasts in bone tumors were determined by immunohistochemistry and the TUNEL assay. Results:In vitro: In vitro treatment with docetaxel inhibited proliferation and resorption pit-forming activity and induced apoptosis of mouse osteoclast cells and UMUC-14 cells. In vitro treatment with minodronate inhibited proliferation and activity and induced apoptosis of mouse osteoclast cells but not UMUC-14 cells. The treatment with minodronate enhanced the inhibition of proliferation and activity by docetaxel in osteoclasts. In vivo: In vivo combination therapy with docetaxel and minodronate significantly reduced the tumor incidence compared with the control (P < 0.05) and also growth of intraossal TCC in athymic nude mice compared with the control (P < 0.001), single therapy with docetaxel (P < 0.01), and minodronate (P < 0.05). Drug-induced body weight loss was not significantly different in any treatment group. Therapy with minodronate significantly enhanced inhibition of proliferation by docetaxel in osteoclasts of bone tumors compared with the control (P < 0.01), single therapy with docetaxel (P < 0.01), and minodronate (P < 0.05). Conclusions: These studies indicate that combination therapy with minodronate and docetaxel may be beneficial in patients with bone metastasis of human TCC in the urinary tract.

A SOLITARY METASTASIS OF RENAL CELL CARCINOMA TO THE URETHRA

A 77-year-old man presented with painless urethral bleeding 1 week in duration. History revealed that 5 years earlier he had undergone right radical nephrectomy for localized pT2N0M0 renal cell carcinoma. Surgical margins were tumor-free on microscopic examination. Histology was consistent with common type, clear cell subtype adenocarcinoma. At 3-year followup chest computerized tomography (CT) revealed 3 3 3 cm. and 2 3 2 cm. lesions in the left lung. However, there were no malignant findings on histological assessment of needle biopsy under CT guidance, and the tumors did not change in size. At 5-year followup the patient had asymptomatic urethral bleeding. Cystoscopy revealed a globular yellow mass on a stalk in the membranous urethra (fig. 1). Metastatic evaluation with CT of the abdomen, excretory urography of the upper urinary tract and bladder, cystoscopy and bone scan was negative. Furthermore, urinary cytology consistently revealed no malignancy. Transurethral resection of the lesion yielded tissue consistent with renal cell carcinoma (fig. 2). Although adjuvant therapies including interferon were recommended, the patient declined further treatment. At 6-month followup the patient was disease-free. He is being followed on an outpatient basis.

Lipoma-like tumor mass probably arising in the retroperitoneal heterotopic pancreas: A previously undescribed lesion

Heterotopic pancreatic tissue is found in several locations of the body. However, to the best of our knowledge, there have been no reports on heterotopic pancreas in the retroperitoneum. A case of retroperitoneal lipoma‐like large tumor mass probably arising in the heterotopic pancreas is reported. A 45‐year‐old Japanese woman was admitted to hospital because of back pain. Imaging modalities showed an abnormal mass in the retroperitoneum separate from the surrounding organs, including the pancreas and kidney. Histologically, the mass consisted of mature adipose tissue and scattered ductal and acinar elements. Although there was no islet tissue in this fatty mass, the epithelial elements suggested heterotopic pancreatic tissue in origin. The present case is very unusual; however, heterotopic pancreas should be considered in the differential diagnosis of retroperitoneal tumors.

IgG4-related disease of the paratestis in a patient with Wells syndrome: a case report

BackgroundWe report a case of a 33-year-old man who presented with immunoglobulin (Ig)G4-related disease (IgG4-RD) forming a pseudotumor in the left paratesticular region during oral administration of corticosteroid for Wells syndrome, which involves cellulitis with eosinophilia.Case presentationThe patient was introduced to our institution from a private hospital with a 3-month history of asymptomatic left scrotal mass. A 5-cm diameter nodule was palpable in the left scrotum. Tumor lesion in the left paratestis involving the epididymis and spermatic cord was observed on computed tomography and magnetic resonance imaging. Blood testing showed no abnormalities other than a minimal increase in C-reactive protein levels. Urine examination likewise revealed no significant findings. Left radical orchidectomy was performed under a diagnosis of left paratesticular neoplasm suspected as malignant tumor. The tumor was pathologically identified as IgG4-RD of the left paratestis involving the epididymis and spermatic cord.ConclusionsWe present a first description of IgG4-RD in a patient with Wells syndrome and the ninth case of IgG4-RD in a scrotal organ, and discuss this very rare entity with reference to the literature.Virtual SlidesThe virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/13000_2014_225

Combination with third-generation bisphosphonate (YM529) and interferon-alpha can inhibit the progression of established bone renal cell carcinoma

The aim of this study was to investigate whether the third‐generation nitrogen‐containing bisphosphonate (YM529) can inhibit the progression of established bone renal cell carcinoma (RCC) and to elucidate its mechanism. Antiproliferative effect and apoptosis induction of RCC cells and mouse osteoclasts by YM529 and/or interferon‐alpha (IFN‐α) were evaluated in vitro using cell counting and in vivo using soft X‐ray, the TUNEL method and tartrate‐resistant acid phosphatase stain. For the in vivo study, male athymic BALB/cA Jc1‐nu nude mice bearing human RCC cell line RBM1‐IT4 cells were treated with YM529 and/or IFN‐α. The biological activity of osteoclasts was evaluated using the pit formation assay. The antiangiogenetic effect by YM529 and/or IFN‐α was analyzed using micro‐vessel density and in situ mRNA hybridization. Osteoclast number in bone tumors was decreased in YM529‐treated mouse. YM529 also inhibited osteoclast activity and proliferation in vitro, whereas basic fibroblast growth factor expressions and micro‐vessel density within tumors were inhibited by IFN‐α. Neither YM529 nor IFN‐α alone significantly inhibited the growth of established bone metastatic tumors. Combined treatment with YM529 and IFN‐α may be beneficial in patients with human RCC bone metastasis. Their effects are mediated by osteoclast recruitment inhibition and inactivation by YM529 and antiangiogenesis by IFN‐α.

Therapy with transcutaneous administration of imiquimod combined with oral administration of sorafenib suppresses renal cell carcinoma growing in an orthotopic mouse model

Imiquimod is an imidazoquinoline immune response modifier that is used in antiviral and antiallergic creams. Combination therapy using transcutaneous imiquimod and oral sorafenib was previously demonstrated to reduce the tumor burden of renal cell carcinoma growing cutaneously in a mouse model. In the present study, an orthotopic mouse model was used to investigate whether combined treatment with oral sorafenib and transcutaneous imiquimod inhibited renal cell carcinoma growing in the kidney. Kidneys of female BALB/c mice were orthotopically implanted with RENCA mouse kidney cancer cells, and the mice were transcutaneously treated with cream containing imiquimod and/or with orally administered sorafenib 5 days following cell implantation. Tumor burden and incidence were determined 28 days following the start of therapy. Splenocyte activity was quantified using the 51Cr release assay and the fluorescence-activated cell sorting assay with cluster of differentiation (CD) 4 and CD8 antibodies. Imiquimod, sorafenib and combination therapy were tolerated well. A combination of transcutaneous imiquimod and oral sorafenib inhibited the growth of RENCA tumors in the kidney significantly compared with the control. The 51Cr release assay demonstrated that transcutaneous imiquimod therapy significantly induced the release of 51Cr from RENCA cells compared with the control. The fluorescence-activated cell sorting assay demonstrated that transcutaneous imiquimod therapy induced CD8+ and CD4- splenocytes compared with the control. In summary, the results of the present study demonstrated that combined treatment with transcutaneous imiquimod and oral sorafenib may be a promising strategy for the treatment of patients with renal cell carcinoma.

低侵襲治療におけるマイクロ波凝固療法,ラジオ波焼灼療法,超音波駆動メスの腎組織に対する影響

PURPOSE The objective of this study was to optimize least invasive techniques, like microwave tissue coagulation (MCT), radiofrequency ablation (RFA) or ultrasonically-activated scalpel (USS) for the electric and thermal conduction in kidney. MATERIALS AND METHODS Needle electrode of MCT, RFA or USS were inserted into pigs kidney, under general anesthesia. We examined if the interruption of renal artery or cooling of renal parenchyma by irrigation in the renal pelvis had an effect on the tissue temperature distribution and area of tissue degeneration. The tissue parenchyma temperatures were measured at regular intervals of area (each 2.5-5.0 mm) from needle electrode or hook probe and of time (each 10 sec) using by needle temperature sensor, and pathological investigations on the tissue degeneration of the renal parenchyma were done. RESULTS By MCT, interruption of renal artery had no effect on the distribution of tissue temperature and the area of tissue degeneration. Cooling of renal parenchyma by irrigation in the renal pelvis did not affect the area of tissue degeneration but the tissue temperature was elevated significantly. By RFA, particular interest, interruption of renal artery and cooling of renal parenchyma enhanced to increase both the tissue temperature and the area of tissue degeneration. By USS, interruption of renal artery enhanced to increase both the tissue temperature and the area of tissue degeneration. Cooling of renal parenchyma by irrigation in the renal pelvis had no effect on the distribution of temperature and the area of tissue degeneration. We also examined weather cooling of renal parenchyma by several irrigation solutions which have different electric conductivity (EC) such as normal saline solution (EC: 15,800 microS/cm), tap water (EC: 133 microS/cm) and glucose solution (EC: 8.6 microS/cm) in the renal pelvis using ureteral catheter would effect on the distribution of temperature and the area of tissue degeneration. In MCT and USS, these solutions did not have any effect, but in RFA the degree of tissue temperature elevation by normal saline, tap water and glucose solution were high, intermediate and low, respectively. CONCLUSION As RFA and USS tend to be affected by the surrounding environment, electric and thermal conductivity should be applied with caution, while using these techniques.