Takeshi Sasaki

Oral Naftopidil Suppresses Human Renal-Cell Carcinoma by Inducing G1 Cell-Cycle Arrest in Tumor and Vascular Endothelial Cells

Renal cell carcinoma (RCC) is an angiogenesis-dependent and hypoxia-driven malignancy. As a result, several targeting agents are being investigated. However, the efficacy of current regimens is generally insufficient for their toxicity and poor overall response rates. We have recently reported that naftopidil exerts growth-inhibitory effects on human prostate cancer cells. In this study, we investigated the biochemical mechanisms by which naftopidil produces growth-inhibitory and antiangiogenic effects on RCC. We first tested the effects of naftopidil on the proliferation of ACHN and Caki-2 RCC cells. Next, we set up a model simulating the tumor microenvironment, in which ACHN cells were grafted onto the renal capsule of mice. We then tested the effects of naftopidil on human umbilical vein endothelial cells cell proliferation and Matrigel plug vascularization. Finally, to establish the antitumor activity of naftopidil on RCC, we tested the antitumor effects of naftopidil on excised tumor specimens from 20 patients with RCC that were grafted beneath the renal capsule of mice. Naftopidil showed similar in vitro growth-inhibitory effects on all cell lines. Fluorescence-activated cell sorting analysis revealed an increase in G1 cell-cycle arrest in all naftopidil-treated cell lines. In vivo tumorigenic studies showed a significant reduction of ACHN tumor weight, Ki-67 index, and microvessel density (MVD) in naftopidil-treated mice. Naftopidil attenuated neovascularization in an in vivo Matrigel plug assay. Studies in mouse xenograft models also showed a significant MVD reduction in naftopidil-treated excised human RCC. The growth-inhibitory effects of naftopidil suggest it may be a novel anticancer agent and a potential preventive option for RCC. Cancer Prev Res; 6(9); 1000–6. ©2013 AACR.

Activation of FGF2-FGFR Signaling in the Castrated Mouse Prostate Stimulates the Proliferation of Basal Epithelial Cells

ABSTRACT The prostate gland is unique in that it undergoes rapid regression following castration but regenerates completely once androgens are replaced. Residual ductal components play an important role in the regeneration of a fully functional prostate. In this study, to examine how androgen status affects prostate structure and components, we conducted histopathological studies of the involuted and regenerated mouse dorsolateral prostate (DLP). In the castrated mouse DLP, the number of luminal epithelial cells decreased in a time-dependent manner. On Day 14 postandrogen replacement, the number of luminal epithelial cells was completely restored to the baseline level. In contrast, the number of basal epithelial cells gradually increased in the castrated mouse prostate. The Ki67-labeling index of prostate basal epithelial cells was significantly increased after castration. The number of basal epithelial cells decreased to baseline after androgen replacement. After castration, mRNA expression levels of specific growth factors, such as Fgf2, Fgf7, Hgf, Tgfa, and Tgfb, were relatively abundant in whole mouse DLPs. In organ culture experiments, basal epithelial proliferation was recapitulated in the absence of dihydrotestosterone (DHT). The proliferation of basal epithelial cells in the absence of DHT was suppressed by treatment with an FGF receptor inhibitor (PD173074). Moreover, FGF2 treatment directly stimulated the proliferation of basal epithelial cells. Taken together, these data indicated that the FGF2-FGF receptor signal cascade in the prostate gland may be one of the pathways stimulating the proliferation of basal epithelial cells in the absence of androgens.

Randomized controlled study of the efficacy and safety of continuous saline bladder irrigation after transurethral resection for the treatment of non-muscle-invasive bladder cancer.

To evaluate the efficacy and safety of continuous saline bladder irrigation (CSBI) after transurethral resection of bladder tumour (TURBT) in patients with low‐ to intermediate‐risk non‐muscle invasive bladder cancer (NMIBC).

Fistula between the external iliac artery and the body of an ileal conduit

Fistulous formation between an ileal conduit and the iliac artery is extremely rare, and it occurrs at the uretero-ileal junction. To our knowledge, this is the first case of a direct fistula between the external iliac artery and the body of an ileal conduit. A 77-year-old man was diagnosed with invasive bladder cancer and underwent radical cystectomy with ileal conduit. Wallace anastomosis was carried out and 6-Fr ureteral stents had been inserted for 8 days after surgery. No leakage of urine was observed when catheters were removed. A total of 51 days after the surgery, stoma bleeding suddenly occurred with right lower quadrant abdominal pain. It stopped spontaneously in a few minutes, but re-bleeding was seen 12 h later. No obvious bleeding point was found by endoscopic examination in the ileal conduit. A total of 60 days after the operation, pulsatile bleeding from the conduit occurred again. Enhanced computed tomography (CT) suggested a pseudoaneurysm of the external iliac artery adjacent to the ileal conduit, although a preoperative CT scan showed no abnormal findings in the right external iliac artery. The existence of a fistula between the external iliac artery and the ileal conduit was strongly suspected (Fig. 1a). Surgical treatment of extra-anatomic arterial reconstruction of the right external iliac artery with common iliac-femoral bypass and excision of the ileal conduit was carried out with cutaneous ureterostomy. Severe adhesion of the ileum was seen around the ileal conduit, and there existed inflammatory change around the right external artery, which might have been a speculated pseudoaneurysm by CT. However, an apparent pseudoaneurysm was not observed (Fig. 1b). The segment of the external iliac artery and ileal conduit containing the fistula was removed en bloc. There was a pinhole fistula in both the cut surface of the external iliac artery and the body of the ileal conduit (Fig. 1c,d); pathological findings revealed the existence of chronic inflammation around the fistula. The patient died of sepsis as a result of peritonitis 1 month after the surgery. Uretero-arterial fistulae can occur in association with prior genitourinary or pelvic surgery, prolonged ureteral stenting, radiation therapy, prior vascular surgery and vascular pathology. The cause of the present case was supposed to be chronic sepsis as a result of minor leakage of urine at uretero-intestinal anastomosis, followed by adhesion and fistula formation. Another factor might be the proximity of the ileal conduit to the external ileac artery, which was rendered vulnerable by being stripped during the en bloc resection of ileac lymph nodes as described by Beaugie et al. They also reported a similar case and noted the presence of a “warning hemorrhage” before the massive bleeding. Recently, there have been several reports of successful treatment for endovascular management of uretero-arterial

Additive naftopidil treatment synergizes docetaxel-induced apoptosis in human prostate cancer cells

AbstractPurposeDocetaxel (DTX) is a standard chemotherapeutic drug for castration-resistant prostate cancer (CRPC), although adverse events are common. To overcome this problem, researchers have evaluated the efficacy of DTX treatment in combination with other drugs. Naftopidil is a tubulin-binding drug with fewer adverse events, implying the usefulness of this drug in clinical applications when combined with DTX. Here, we investigated the efficacy of additive naftopidil treatment in combination with DTX on prostate cancer (PCa) cells.nMethodsThe effects of combination treatment with DTX plus naftopidil were analyzed using two animal models of LNCaP cells plus PrSC xenografts (sub-renal capsule grafting) and PC-3 xenografts (intratibial injection).ResultsCombination treatment with DTX plus naftopidil significantly inhibited cell growth in LNCaP cells compared with DTX alone. Analysis of the cooperativity index (CI) showed that combination treatment exhibited additive effects on DTX-induced growth inhibition in LNCaP cells. In contrast, combination treatment showed more than an additive (synergistic) effect on DTX-induced apoptosis in LNCaP and PC-3 cells. In LNCaP cells plus PrSC xenografts, combination treatment showed synergistic effects on DTX-induced apoptosis. The synergistic effects of naftopidil on DTX-induced apoptosis were also observed in PC-3 xenografts.ConclusionsOur results demonstrated that additive naftopidil treatment in combination with DTX increased the efficacy of DTX for the treatment of LNCaP and PC-3 tumors in vivo. Thus, additive naftopidil treatment showed a synergistic effect on DTX-induced apoptosis in PCa cells in vitro and in vivo, suggesting that this treatment approach may yield improved clinical benefits compared with DTX alone.

Predicting the tumorigenic phenotype of human bladder cancer cells by combining with fetal rat mesenchyme

BACKGROUNDnIn nonmuscle invasive bladder cancer patients, prediction of pTa and pT1 bladder cancer recurrence and progression must be established. Micropapillary structures have been defined as small clusters of invasive cancer cells having features of the epithelial-mesenchymal transition. Since the stromal microenvironment helps to induce the epithelial-mesenchymal transition, interactions between cancer cells and stroma should be closely examined to predict the tumorigenic phenotype of human bladder cancer cells.nnnMATERIALS AND METHODSnTo investigate differences in the responsiveness of cancer cells to stroma, we combined 3 established human bladder cancer cell lines (high-grade T24 and UM-UC-3 cells, and low-grade papillary RT4 cells) with fetal rat mesenchyme.nnnRESULTSnAmong 3 bladder cancer cell lines, the expression profiles of p63 isoforms were distinct, i.e., p63γ in T24 cells, p63β in UM-UC-3 cells, and p63α in RT4 cells. Tumors formed by T24 cells combined with fetal mesenchyme formed micropapillary-like structures, whereas those formed by T24 cells alone did not. T24 cells combined with fetal mesenchyme showed poor differentiation, e.g., innumerable chromatic atypia in the nuclei, higher levels of chromatic condensation, and increased nucleoli. In contrast, both UM-UC-3 and RT4 cells combined with fetal mesenchyme did not form micropapillary-like structures. Ki-67 and p63 labeling indices were significantly elevated by combining fetal mesenchyme with T24 cells but not with the others.nnnCONCLUSIONSnBy mixing cancer cells with fetal mesenchyme, our data demonstrated that formation of micropapillary-like structures may predict the tumorigenic phenotype of invasive bladder cancer cells. Taken together, distinct expression profiles of p63 isoforms may predict poor outcomes in invasive bladder cancer.

Interleukin-6 induces VEGF secretion from prostate cancer cells in a manner independent of androgen receptor activation

The reduced androgen‐sensitivity of prostate cancer (PCa) cells is an important clinical development because of its association with the cells’ progression to castration‐resistant prostate cancer (CRPC). During androgen deprivation therapy (ADT), stroma‐derived growth factors and cytokines can activate the androgen receptor (AR). For example, IL‐6 is a multifunctional cytokine that is involved in the malignancy of PCa cells through AR activation. In the present study, we used an androgen‐sensitive human PCa cell line (LNCaP) and its sublines to investigate the relationship between the responsiveness of PCa cells to IL‐6 treatment and the cellular AR signaling pathway.

Successful treatment of non‐invasive bladder tumour in a haemophilia A patient with high‐responding inhibitors: a case report

T. SASAKI,* T. MATSUMOTO,†‡ H. WADA,§ K. OHISHI,†‡ H. KANDA,* Y. YAMADA,* K. ARIMA,* N. KATAYAMA‡ and Y. SUGIMURA* *Department of Nephro-Urologic Surgery and Andrology, Mie University Graduate School of Medicine; †Division of Blood Transfusion, Mie University Hospital; ‡Department of Haematology; and §Department of Molecular and Laboratory Medicine, Mie University Graduate School of Medicine, Tsu, Mie, Japan

1649 THE BENEFIT OF CONTINUOUS SALINE BLADDER IRRIGATION AFTER TRANSURETHRAL RESECTION IN NON-MUSCULAR INVASIVE BLADDER CANCER

non-muscle invasive bladder cancer (NMIBC). A prospective, long term, randomized comparison with a similar series initially diagnosed by white light cystoscopy (WLC) alone with regard to recurrence rates was also performed. METHODS: A total of 362 patients suspected of NMIBC were included in the trial, based on positive urinary cytology and ultrasonographic suspicion of bladder tumors. The 181 patients of the BL arm underwent both WLC and BLC. A single postoperative mytomicin-C instillation was performed in all cases, intravesical chemotherapy for intermediate risk patients and BCG instillations for high-risk cases. The follow-up protocol consisting of urinary cytology and WLC was also performed in accordance with the risk category of NMIBC cases for a period of two years. Only first time recurrences after the initial diagnostic were taken into consideration. RESULTS: A total of 142 NMIBC patients were diagnosed in the BL arm and 129 in the WL arm. In the BL series, the CIS, pTa, pT1 and overall cases’ and tumors’ detection rates were significantly improved for BLC. Additional tumors were found by WLC and BLC in 14.1% versus 35.2% of the cases. Consequently, the recurrence and progression risk categories of patients changed in 4.9% and 7% of the cases due to WLC and in 16.2% and 21.1% due to BLC additionally found tumors. The postoperative treatment changed in 6.3% of the cases due to WLC and in 19% due to BLC results. A total of 125 patients of the BL group and 114 of the WL group completed the follow-up protocol. The recurrence rate at 3 months was 15.8% in the WL series versus 7.2% in the BL series. The rates of primary site and other site recurrences in the WL and BL arms were 7.9% versus 5.6% and 6.1% versus 0.8%. The one and two years’ recurrence rates were significantly higher in the WLC group by comparison to the BLC group (32.5% versus 21.6% and 45.6% versus 31.2%). CONCLUSIONS: HAL-BLC cystoscopy emphasized superior patients’ and tumors’ detection rates in NMIBC cases, as well as a significant impact in terms of risk category changes and consequent postoperative treatment modifications. At the 3 months’ follow-up, significantly improved recurrence rates were determined in the BL arm, mostly due to fewer other site recurrences. The one and two years’ recurrence rates confirmed the long term advantages of HAL-BLC.