Hideaki Kise

Purification and characterization of a novel calcium-binding protein, S100C, from porcine heart

A novel Ca(2+)-binding protein, which we have named S100C (Ohta et al. (1991) FEBS Lett. 295, 93-96), was purified to homogeneity from porcine heart by Ca(2+)-dependent dye-affinity chromatography. S100C possesses some properties of S100 proteins, such as self-association and exposure of a hydrophobic site upon binding of Ca2+ but it differs from S100 proteins in forms of its isoelectric point (pI = 6.2), cross-reactivity with antibodies, staining by Stains-all, and its Ca(2+)-dependent interaction with the immobilized dye. S100C bound to cytoskeletal components at physiological concentrations of Ca2+. Moreover, it was found that 125I-labeled S100C interacted with annexin I in a Ca(2+)-dependent manner. S100C also inhibited the phosphorylation of annexin I by protein kinase C. These data suggest that S100C might act to regulate the cytoskeleton in a Ca(2+)-dependent manner via interactions with annexin I.

Naftopidil, a Selective α1-Adrenoceptor Antagonist, Suppresses Human Prostate Tumor Growth by Altering Interactions between Tumor Cells and Stroma

In prostate cancer, tumor–stroma interactions play a critical role in the promotion of tumorigenesis, and thus the prevention of those interactions is a promising target to suppress tumor growth. Several studies demonstrated that alpha1-adrenoceptor (α1-AR) antagonists, therapeutic drugs for benign prostatic hyperplasia, have growth inhibitory effects on human prostate cancer (PCa) cells through induction of apoptosis or G1 cell-cycle arrest. However, their direct actions on stromal cells surrounding cancer cells have not yet been elucidated. In this study, we investigated the effects of subtype-selective α1-AR antagonists (naftopidil, tamsulosin, and silodosin) on prostate tumor growth with a focus on the role of stroma, using commercially available fibroblast cells (PrSC). Tumorigenic studies in vivo showed significant reductions in tumor growth when E9 cells (an androgen low-sensitive LNCaP subline) grafted with PrSC were treated with naftopidil. In in vitro analyses, naftopidil and silodosin showed antiproliferative effects on PCa cells regardless of androgen sensitivity and α1-AR subtype expression. In PrSC, a strong growth inhibitory effect was observed with naftopidil but not silodosin. Flow cytometric analysis revealed that naftopidil, but not silodosin, induced G1 cell-cycle arrest in both PCa cells and PrSC. In naftopidil-treated PrSC, total interleukin-6 protein was significantly reduced with increased suppression of cell proliferation. Silodosin induced weak early apoptosis only in PCa cells. These findings demonstrated that naftopidil strongly suppressed cell proliferation of stromal cells, resulting in decreased tumorigenic soluble factor, suggesting that naftopidil might be effective in preventing stromal support of tumor cells. Cancer Prev Res; 4(1); 87–96. ©2011 AACR.

Intermittent docetaxel therapy with estramustine for hormone-refractory prostate cancer in Japanese patients.

BackgroundWe evaluated the efficacy and toxicity of intermittent docetaxel (DCT) with estramustine (EM) for hormone-refractory prostate cancer (HRPC).MethodsFifteen patients were enrolled. They received injected DCT (70 mg/m2 body surface) on day 1 in association with oral EM 560 mg/day (days 1–5). Treatments were repeated every 3 weeks. Serum prostate-specific antigen (PSA) levels were categorized based on the first three courses. Patients exhibiting either a response or stable disease (SD) could have a holiday from treatment (intermittent schedule). The holiday continued until elevation of the PSA level from the nadir baseline level occurred three times. All patients were evaluated for toxicity and quality of life (QOL). Survival curves were established using Kaplan-Meier graphs.ResultsThe median number of courses of DCT/EM therapy was five (range, 3–12 courses). The response rate of the first cycle was 53%: 3 patients with complete response (CR), 5 patients with partial response (PR), 4 patients with SD, and 3 patients with disease progression. Eight patients were able to begin the second re-entry cycle. No patients showed a CR, 2 patients exhibited PR, 4 patients had SD, and the overall response rate was 25%. The survival rates were 93% at 1 year, and 26.1% at 2 years Grade 3–4 anemia was observed in 2 patients (13.3%), neutropenia in 11 (73.3%), and thrombocytopenia in 2 (13.3%). The QOL scale showed good QOL after 6 months, with improvement in the score for nausea and vomiting.ConclusionIntermittent DCT/EM therapy was well tolerated, and has the potential to prolong survival, with a high QOL, in patients with HRPC.

Comparison of radical nephrectomy techniques in one center: Minimal incision portless endoscopic surgery versus laparoscopic surgery

This study was designed to assess the intraoperative and postoperative benefits of two techniques for treating renal cell carcinoma (portless endoscopic surgery with radical nephrectomy [PLES‐RN] and laparoscopic radical nephrectomy [LRN]) carried out at a single center. Radical nephrectomy with either PLES‐RN (14 cases) or LRN (15 cases) was carried out on 29 patients with cT1 renal cell carcinoma. There were no statistically significant between‐group differences in patient characteristics (except tumor side), operation time, and amount of blood loss (χ2 and Fishers test). No blood transfusions were required in either group. The mean incision length of PLES‐RN was not significantly longer than that of LRN. No minor or major complications resulted. From postoperative data, the first intake of fluid (P = 0.07) and food (P = 0.02) tended to be sooner in the PLES group than the LRN group. Postsurgically, white blood cell count and C‐reactive protein were not significantly different between the two groups. The added cost of disposable instruments needed in LRN was 111 570 Japanese yen (1115.7 United States dollars). Both techniques are optimal options for surgically treating early renal cell carcinoma. The comparison related to invasiveness between the two methods should be evaluated using a large number of cases focusing on the various aspects for the future.

Improvement in predicting tumorigenic phenotype of androgen-insensitive human LNCaP prostatic cancer cell subline in recombination with rat urogenital sinus mesenchyme

Hormone‐refractory prostate cancer, a heterogeneous disease, has varying degrees of androgen sensitivity. To understand the physiological changes in the hormone‐refractory state, the present study used a lineage‐derived androgen receptor (AR)‐positive, androgen‐insensitive prostate cancer cell line and evaluated the tumorigenic phenotype, focusing on tumor–stromal interactions in vivo. First, tumorigenic differences of cancer cells alone were examined in an androgen‐insensitive AR‐positive LNCaP subline, AIDL, compared with those of the androgen‐sensitive AR‐positive parental LNCaP and the androgen‐insensitive AR‐negative PC‐3 cells transplanted into subcutaneous, sub‐renal and prostatic orthotopic graft sites. Next, cancer cells were recombined with rat urogenital sinus mesenchyme (rUGM) to simulate the tumor‐stromal microenvironment. Tumors of AIDL and LNCaP without stromal components both formed well‐defined globular tumors and contained large blood‐filled areas, with no significant difference in tumor growth or histopathology regardless of the cell lines androgen sensitivity or graft site. In contrast, tumors of AIDL and LNCaP recombined with rUGM both showed reduction of blood‐filled areas in the tumors and increased tumor growth compared with cancer cells alone. Tumors of AIDL + rUGM recombinants were approximately three times as large as those of LNCaP + rUGM recombinants, whereas tumors of AIDL and LNCaP without rUGM were not different in size. In addition to the tumor size, cell proliferation (Ki‐67 labeling index) in tumors of AIDL + rUGM recombinants was significantly higher than that in tumors of LNCaP + rUGM recombinants. Immunoreactivities of AR, E‐cadherin and β‐catenin were decreased in AIDL + rUGM recombinants relative to AIDL alone and LNCaP + rUGM recombinants. These results demonstrated that tumorigenic features of androgen‐insensitive AR‐positive prostate cancer cells could be significantly influenced by rUGM. Therefore, this in vivo recombination model with rUGM may be useful in developing new treatment strategies. (Cancer Sci 2008; 99: 2435–2443)

Paravesical granuloma after inguinal herniorrhaphy. Case report and review of the literature.

Paravesical granuloma after herniorrhaphy is an unusual complication due to infected suture material and often mimics bladder or urachal malignancy. We present 3 cases of this disease and reviewed 21 previously reported cases. Our patients underwent hernial repair 2–7 years before examination and presented urinary symptoms. They were treated with antibiotics and underwent exploration of the inguinal wound infection but the symptoms did not resolve. En bloc excision and partial cystectomy were performed and resulted in complete resolution of the symptoms. It is important to consider paravesical granuloma in patients who had undergone herniorrhaphy in the differential diagnosis of bladder or urachal tumors.

Gemcitabine and capecitabine chemotherapy in Japanese patients with immunotherapy‐resistant renal cell carcinoma

The objective of this study was to evaluate the efficacy and toxicity of combined gemcitabine and capecitabine (Gca) chemotherapy in patients with advanced renal cell cancer after immunotherapy failure. Nine patients were enrolled in this trial. Gemcitabine (1000 mg/m2) was injected on days 1 and 8, followed by oral administration of capecitabine (1660 mg/m2) on days 1–14. The response rate was 11%, with a partial response in one patient (11%), stable disease in five patients (56%) and disease progression in three patients (33%). Grade 3–4 neutropenia was observed in one patient (11%) and thrombocytopenia in two patients (22%). The quality of life (QOL) questionnaire scales showed no significant changes induced by chemotherapy. The median progression‐free survival was 4 months with an overall 1‐year survival rate of 78%. Gemcitabine and capecitabine chemotherapy can be safely administered as second‐line therapy in renal cell cancer patients, maintaining QOL baseline parameters.

α-Fetoprotein producing urachal tumor

A 41-year-old woman was admitted to hospital with a 3-month history of urinary frequency and macroscopic hematuria. Magnetic resonance imaging demonstrated a cystic mass extending to the urachus, which compressed the anterior wall of the bladder. Cystoscopy showed a nonpapillary tumor at the dome of the bladder and biopsy revealed anaplastic adenocarcinoma. Serum AFP was 23,226 ng./ml. (normal less than 28.5) and serum carcinoembryonic antigen was 20.4 ng./ml. (normal less than 6). Human chorionic gonadotropin was normal. Gastroduodenoscopy and barium enema revealed no malignancy. Diagnosis was urachal tumor derived from a cyst with invasion into the bladder. En bloc resection of the tumor, including the umbilicus, urachus, peritoneum and bladder dome, was performed. The cystic mass, which contained yellowish fluid, had a thick hard wall invading to the dome of the bladder (fig. 1). On histological examination the tumor was poorly differentiated tubular adenocarcinoma, and was immunoreactive for AFP and carcinoembryonic antigen (fig. 2). There were no histological features of hepatoid carcinoma. Serum AFP and carcinoembryonic antigen decreased to normal ranges within 2 months postoperatively.

Effect of transforming growth factor α overexpression on urogenital organ development in mouse.

Transforming growth factor-α (TGFα) promotes cell proliferation by binding to the epidermal growth factor receptor (EGFR). TGFα and EGFR overexpression have been reported in various human cancers. However, whether TGFα induces cancer by itself is unknown in urogenital organs. To investigate whether TGFα overexpression induces carcinogenesis in urogenital organs, we analyzed the phenotypes of urogenital organs in male TGFα transgenic (TG) mice of the CD1 strain. Urogenital organs including the kidney, bladder, prostate, seminal vesicles, testes, and epididymis were isolated from 4- to 48-week-old TGFα TG and wild-type (WT) CD1 mice. Prostates were separated into anterior prostate (AP), dorsolateral prostate (DLP), and ventral prostate (VP). Neither tumor formation nor epithelial hyperplasia was observed in the TGFα TG mouse urogenital organs that we have investigated. Histopathologically, in prostate, we found an increased number of p63-positive basal epithelial cells in the TGFα TG mice AP and DLP. There was no morphological change in the stromal component, such as hypercellular stroma or fibrosis. However, bladder weight was greater in TGFα TG mice than that in WT mice, and distended bladders were observed macroscopically in 19 of 20 TGFα TG mice over 20 weeks of age. Ki67 labeling index was increased significantly in the TGFα TG mouse urethral epithelium, whereas neither epithelial hyperplasia nor hypertrophy was observed. In conclusion, our results suggest that TGFα overexpression in mouse urogenital organs alone may not be responsible for tumor formation and epithelial hyperplasia, but is involved in bladder outlet obstruction.

Manserin as a novel histochemical neuroendocrine marker in prostate cancer.

OBJECTIVES To investigate the presence of manserin in human prostate cancers and to correlate manserin expression with pathologic outcomes and progression-free survival. METHODS Eighty-seven patients with recent prostate cancer were classified into 4 groups based on Gleason score, and manserin immunohistochemistry was correlated with Gleason sum grade. To investigate the validity of manserin as a prognostic factor, the Cox proportional hazards regression model was performed on 48 patients in our cohort with T3 or T4 prostate cancer who were initially treated with androgen deprivation therapy. RESULTS The manserin-positive rates of patients with Gleason sums of 6, 7, 8, and ≥9 were 0%, 20.0%, 35.0%, and 48.1%, respectively. Manserin-positive rates were positively correlated with Gleason sums (P = 0.0001). Median times to cancer progression in groups with (n = 8) and without (n = 40) manserin expression were 8 months and 28 months, respectively (P = 0.01). Univariate Cox analysis revealed that manserin expression, clinical stage T4, and high Gleason sum were significantly associated with progression. Multivariate analysis revealed that only 2 factors, manserin expression (hazard ratio (HR) 4.99, P = 0.01) and clinical stage T4 (HR 4.77, P = 0.03), were independent risk factors for progression. CONCLUSIONS This is the first report of manserin expression in human prostate cancers. Manserin may serve as a marker of prostate cancer progression.