Sachiyo Nishida

Prognostic impact of the expression of ALDH1 and SOX2 in urothelial cancer of the upper urinary tract

Aldehyde dehydrogenase 1 (ALDH1) and sex determining region-Y-related high mobility group box 2 (SOX2) have been identified as putative cancer stem-like cell/tumor-initiating cell markers in various cancer tissues. The aim of this study was to elucidate the prognostic impact of these putative cancer stem-like cell/tumor-initiating cell markers in upper urinary tract urothelial cell carcinoma. Immunohistochemical staining for ALDH1 and SOX2 was carried out on archival specimens from 125 patients with upper urinary tract urothelial cell carcinoma who underwent radical nephroureterectomy. The prognostic value of ALDH1 and SOX2 expression and other clinicopathological features was evaluated. On univariate analysis, tumor grade, pathological T stage, pathological N stage, lymphovascular invasion, ALDH1 expression and SOX2 expression were associated with a poor prognosis. On multivariate analysis, the independent factors of prognosis were tumor grade (P=0.014), pathological N stage (P=0.005) and ALDH1 expression (P=0.002). In subgroup analysis, those subgroups with no positive, one positive or two positive results in immunohistochemistry for ALDH1 and SOX2 expression had estimated 5-year cancer-specific survival rates of 80%, 49% and 22%, respectively (P<0.001). Neither ALDH1 nor SOX2 expression correlated with intravesical recurrence after radical nephroureterectomy. These findings suggest that cancer stem-like cells/tumor-initiating cells are linked to more aggressive behavior of upper urinary tract urothelial cell carcinoma, supporting the current cancer stem cell hypothesis. Thus, therapeutic targeting of cancer stem-like cells/tumor-initiating cells in upper urinary tract urothelial cell carcinoma is a future possibility.

Prostate cancer stem-like cells/cancer-initiating cells have an autocrine system of hepatocyte growth factor.

Prostate cancer cells include a small population of cancer stem‐like cells (CSCs)/cancer‐initiating cells (CICs) that have roles in initiation and progression of the cancer. Recently, we isolated prostate CSCs/CICs as aldehyde dehydrogenase 1‐highh (ALDH1high) cells using the ALDEFLUOR assay; however, the molecular mechanisms of prostate CSCs/CICs are still elusive. Prostate CSCs/CICs were isolated as ALDH1high cells using the ALDEFLUOR assay, and the gene expression profiles were analyzed using a cDNA microarray and RT‐PCR. We found that prostate CSCs/CICs expressed higher levels of growth factors including hepatocyte growth factor (HGF). Hepatocyte growth factor protein expression was confirmed by enzyme linked immunosorbent assay and Western blotting. On the other hand, c‐MET HGF receptor was expressed in both CSCs/CICs and non‐CSCs/CICs at similar levels. Hepatocyte growth factor and the supernatant of myofibloblasts derived from the prostate augmented prostasphere formation in vitro, and prostasphere formation was inhibited by an anti‐HGF antibody. Furthermore, c‐MET gene knockdown by siRNA inhibited the prostasphere‐forming ability in vitro and tumor‐initiating ability in vivo. Taken together, the results indicate that HGF secreted by prostate CSCs/CICs and prostate myofibroblasts has a role in the maintenance of prostate CSCs/CICs in an autocrine and paracrine fashion.

Gene expression profiles of prostate cancer stem cells isolated by aldehyde dehydrogenase activity assay.

PURPOSE Prostate cancer cells include a small population of cancer stem-like/cancer initiating cells, which have roles in cancer initiation and progression. Recently aldehyde dehydrogenase activity was used to isolate stem cells of various cancer and normal cells. We evaluated the aldehyde dehydrogenase activity of the human prostate cancer cell line 22Rv1 (ATCC®) with the ALDEFLUOR® assay and determined its potency as prostate cancer stem-like/cancer initiating cells. MATERIALS AND METHODS The human prostate cancer cell line 22Rv1 was labeled with ALDEFLUOR reagent and analyzed by flow cytometry. ALDH1(high) and ALDH1(low) cells were isolated and tumorigenicity was evaluated by xenograft transplantation into NOD/SCID mice. Tumor sphere forming ability was evaluated by culturing in a floating condition. Invasion capability was evaluated by the Matrigel™ invasion assay. Gene expression profiling was assessed by microarrays and reverse transcriptase-polymerase chain reaction. RESULTS ALDH1(high) cells were detected in 6.8% of 22Rv1 cells, which showed significantly higher tumorigenicity than ALDH1(low) cells in NOD/SCID mice (p < 0.05). Gene expression profiling revealed higher expression of the stem cell related genes PROM1 and NKX3-1 in ALDH1(high) cells than in ALDH1(low) cells. ALDH1(high) cells also showed higher invasive capability and sphere forming capability than ALDH1(low) cells. CONCLUSIONS Results indicate that cancer stem-like/cancer initiating cells are enriched in the ALDH1(high) population of the prostate cancer cell line 22Rv1. This approach may provide a breakthrough to further clarify prostate cancer stem-like/cancer initiating cells. To our knowledge this is the first report of cancer stem-like/cancer initiating cells of 22Rv1 using the aldehyde dehydrogenase activity assay.

Potential Survival Benefit of Anti-Apoptosis Protein: Survivin-Derived Peptide Vaccine with and without Interferon Alpha Therapy for Patients with Advanced or Recurrent Urothelial Cancer—Results from Phase I Clinical Trials

We previously identified a human leukocyte antigen (HLA)-A24-restricted antigenic peptide, survivin-2B80–88, a member of the inhibitor of apoptosis protein family, recognized by CD8+cytotoxic T lymphocytes (CTL). In a phase I clinical trial of survivin-2B80-88 vaccination for metastatic urothelial cancer (MUC), we achieved clinical and immunological responses with safety. Moreover, our previous study indicated that interferon alpha (IFNα) enhanced the effects of the vaccine for colorectal cancer. Therefore, we started a new phase I clinical trial of survivin-2B80–88 vaccination with IFNα for MUC patients. Twenty-one patients were enrolled and no severe adverse event was observed. HLA-A24/survivin-2B80–88 tetramer analysis and ELISPOT assay revealed a significant increase in the frequency of the peptide-specific CTLs after vaccination in nine patients. Six patients had stable disease. The effects of IFNα on the vaccination were unclear for MUC. Throughout two trials, 30 MUO patients received survivin-2B80–88 vaccination. Patients receiving the vaccination had significantly better overall survival than a comparable control group of MUO patients without vaccination (P = 0.0009). Survivin-2B80–88 vaccination may be a promising therapy for selected patients with MUC refractory to standard chemotherapy. This trial was registered with UMIN00005859.

Prostate cancer detection by prebiopsy 3.0-Tesla magnetic resonance imaging

Objectives:  The diagnostic value of 3.0‐Tesla magnetic resonance imaging (MRI) for prostate cancer remains to be determined. The aim of the present study was to assess the features of prostate cancer detectable by prebiopsy 3.0‐Tesla MRI.

Influence of Body Mass Index and Total Testosterone Level on Biochemical Recurrence Following Radical Prostatectomy

OBJECTIVE A high body mass index (BMI) and a low testosterone level were recently reported to be prognostic factors for prostate-specific antigen (PSA) recurrence following radical prostatectomy (RP). The goal of this study was to clarify their relationship and influences on biochemical recurrence after RP. METHODS We analysed 126 patients whose data, including the pre-operative BMI and pre-operative serum total testosterone level, were available. All patients underwent RP at our institution between March 1998 and April 2006 without any adjuvant therapy or pelvic lymph node metastasis. The Cox proportional hazards model was used for the multivariate analysis regarding PSA recurrence for the variables of age, operation period, BMI, clinical stage, PSA, Gleasons sum, pre-operative serum total testosterone level and margin status. RESULTS There were no internal correlations among the parameters we used, even between BMI and the total testosterone level. The total testosterone level was not different between two BMI groups (BMI <26.4 and >/=26.4 kg/m(2): the cut-off is the mean + 1 SD). BMI, PSA and Gleasons sum were found to be independent predictors for PSA recurrence through the multivariate analysis. PSA recurrence-free survival rates at 2 years were 77% for BMI <26.4 kg/m(2), and 31% for BMI >/=26.4 kg/m(2) (P = 0.002, log-rank test, 95% CI: 1.489-7.726). CONCLUSIONS The current study suggests that high BMI independently contributes to PSA recurrence but that the total testosterone level does not. Although the mechanism by which obesity promotes PSA recurrence in RP patients has not been established, careful observation is needed for patients with high BMI.

Rectal injury during laparoscopic radical prostatectomy: detection and management.

Among 294 patients who underwent laparoscopic radical prostatectomy (LRP), five (1.7%) developed complications such as rectal injury (RI) and rectourethral fistula (RUF). In four patients, the RI was immediately repaired by placing two layers of uninterrupted sutures without fecal diversion. The RI in two of these four patients were diagnosed using a transrectally inserted Hegar uterine dilator (26 mm). The remaining patients, who presented with RUF as the primary manifestation, were conservatively managed, and the fistulas closed spontaneously. Most of the RI detected during the operation were managed with primary fistula closure without fecal diversion. In some cases of postoperative RUF, spontaneous closure may occur while the patient is waiting for surgical repair.

New technique with combination of felt, Hem-o-lok and Lapra-Ty for suturing the renal parenchyma in laparoscopic partial nephrectomy

We reported a new technique for closure of the renal parenchyma in laparoscopic partial nephrectomy, shortening the suturing time. Between 2009 and 2011, 41 patients with renal masses 4 cm or smaller in diameter underwent transabdominal laparoscopic partial nephrectomy by a single surgeon in a single institution. The sutures were carried out using 2‐0 vicryl CT‐1 with a 1.2 × 1.2 cm piece of felt, and both sutures were temporarily held using a Hem‐o‐lok. After all sutures (median 3) were completed, they were sequentially fixed by sliding the Hem‐o‐lok, and then locked using the Lapra‐Ty. The median times for suturing the renal parenchyma and ischemic time were 13 min and 28 min, respectively. The arrangement of the wound and hemostasis were good. No patients developed urinoma or postoperative bleeding.

Adrenocortical carcinoma : Retrospective study of 14 patients experienced at a single institution over 34 years

Objective:  To review clinical outcome of patients with adrenocortical carcinoma experienced at a single institute over 34 years.

Lymph node metastasis mapping in extended lymphadenectomy to the level of the inferior mesenteric artery for bladder cancer

BackgroundThe aim of this study was to evaluate the distribution of lymph node metastasis in extended lymphadenectomy for patients with bladder cancer.MethodsWe analyzed 31 patients who underwent extended lymphadenectomy at radical cystectomy for bladder cancer between April 2008 and February 2010. Specimens were evaluated as 14 separate packages from predesignated anatomical locations. The lymph node mapping was prospectively registered.ResultsThe median lymph node count was 37 (range 19–68). Ten (32%) patients had lymph node metastasis. The positive rates at each lymph node site were 0% at the left internal iliac, 13% at the left obturator, 3.2% at the left external iliac, 6.5% at the right internal iliac, 10% at the right obturator, 16% at the right external iliac, 3.2% at the left common iliac, 3.2% at the right common iliac and 6.5% at the presacral node. No lymph node metastasis was detected in the Cloquet, paracaval, aortocaval or paraaortic nodes. One (3.2%) patient had a skip metastasis from the left obturator to the presacral node.ConclusionsExtended lymphadenectomy provides more accurate lymph node staging. We suggest that it is better to perform lymphadenectomy at least below the aortic bifurcation including the presacral node.