XAF1 expression and regulatory effects of somatostatin on XAF1 in prostate cancer cells

<p>Abstract</p> <p>Background</p> <p>Somatostatin prevents cell proliferation by inducing apoptosis. Downregulation of the <it>XAF1 </it>transcript may occur during the development of prostate cancer. It is interesting to evaluate the potential regulatory effects of somatostatin on <it>XAF1 </it>expression during the development of prostate cancer cells.</p> <p>Methods</p> <p><it>XAF1 </it>mRNA and protein expression in human prostate epithelial cells RWPE-1, androgen dependent prostate cancer LNCaP, and androgen independent DU145 and PC3 cells were evaluated using RT-PCR and Western blot. The regulation of <it>XAF1 </it>mRNA and protein expression by somatostatin and its analogue Octreotide was evaluated.</p> <p>Results</p> <p>Substantial levels of <it>XAF1 </it>mRNA and proteins were detected in RWPE-1 cells, whereas prostate cancer cells LNCaP, DU145 and PC3 exhibited lower <it>XAF1 </it>expression. Somatostatin and Octreotide up-regulated <it>XAF1 </it>mRNA and protein expression in all prostate cancer cell lines.</p> <p>Conclusions</p> <p><it>XAF1 </it>down-regulation may contribute to the prostate cancer development. The enhanced <it>XAF1 </it>expression by somatostatin indicates a promising strategy for prostate cancer therapy.</p

Urethral Reconstruction Using Mesothelial Cell-Seeded Autogenous Granulation Tissue Tube: An Experimental Study in Male Rabbits

Objective. This study was to evaluate the utility of the compound graft for tubularized urethroplasty by seeding mesothelial cells onto autogenous granulation tissue. Methods. Silastic tubes were implanted subcutaneously in 18 male rabbits, of which nine underwent omentum biopsies simultaneously for in vitro expansion of mesothelial cells. The granulation tissue covering the tubes was harvested 2 weeks after operation. Mesothelial cells were seeded onto and cocultured with the tissue for 7 days. A pendulous urethral segment of 1.5 cm was totally excised. Urethroplasty was performed with mesothelial cell-seeded tissue tubes in an end-to-end fashion in nine rabbits and with unseeded grafts in others as controls. Serial urethrograms were performed at 1, 2, and 6 months postoperatively. Meanwhile, the neourethra was harvested and analyzed grossly and histologically. Results. Urethrograms showed cell-seeded grafts maintained wide at each time point, while strictures formation was found in unseeded grafts. Histologically, layers of urothelium surrounded by increasingly organized smooth muscles were observed in seeded grafts. In contrast, myofibroblasts accumulation and extensive scarring occurred in unseeded grafts. Conclusions. Mesothelial cell-seeded granulation tissue tube can be successfully used for tubularized urethroplasty in male rabbits

Simvastatin inhibits renal cancer cell growth and metastasis via AKT/mTOR, ERK and JAK2/STAT3 pathway.

Renal cell carcinoma (RCC) is the most lethal type of genitourinary cancer due to its occult onset and resistance to chemotherapy and radiation. Recently, accumulating evidence has suggested stains, inhibitors of 3-hydroxy-3-methyl glutaryl coenzyme A (HMG-CoA) reductase, were associated with the risk reduction of cancer. In the present study, we aimed to investigate the potential effects of simvastatin on RCC cells and the underlying mechanisms by which simvastatin exerted its actions. With cell viability, colony formation, and flow cytometric apoptosis assays, we found that simvastatin potently suppressed cell growth of A498 and 786-O cells in a time- and dose- dependent manner. Consistently, the xenograft model performed in nude mice exhibited reduced tumor growth with simvastatin treatment. In addition, the inhibitory effects of simvastatin on migration and invasion were also observed in vitro. Mechanically, we presented that simvastatin could suppress the proliferation and motility of RCC cells via inhibiting the phosphorylation of AKT, mTOR, and ERK in a time- and dose- dependent manner. Further investigation of the underlying mechanism revealed simvastatin could exert the anti-tumor effects by suppressing IL-6-induced phosphorylation of JAK2 and STAT3. In conclusion, these findings suggested that simvastatin-induced apoptosis and its anti-metastasis activity in RCC cells were accompanied by inhibition of AKT/mTOR, ERK, and JAK2/STAT3 pathways, which imply that simvastatin may be a potential therapeutic agent for the treatment of RCC patients

Pretreatment Serum Cystatin C Levels Predict Renal Function, but Not Tumor Characteristics, in Patients with Prostate Neoplasia

To evaluate the role of Cystatin C (Cys-C) in tumorigenesis and progression of prostate cancer (PCa), we retrospectively collected the clinical information from the records of 492 benign prostatic hyperplasia (BPH), 48 prostatic intraepithelial neoplasia (PIN), and 173 PCa patients, whose disease was newly diagnosed and histologically confirmed. Pretreatment serum Cys-C levels were compared across the various groups and then analyzed to identify relationships, if any, with clinical and pathological characteristics of the PCa patient group. There were no significant differences in serum Cys-C levels among the three groups (P > 0.05). In PCa patients with normal SCr levels, patient age was correlated with serum Cys-C level (P ≤ 0.001) but did not correlate with alkaline phosphatase (AKP), lactate dehydrogenase (LDH), prostate specific antigen (PSA), Gleason score, or bone metastasis status (P > 0.05). Age and SCr contributed in part to the variations in serum Cys-C levels of PCa patients (r = 0.356, P ≤ 0.001; r = 0.520, P ≤ 0.001). In conclusion, serum Cys-C levels predict renal function in patients with prostate neoplasia, but were not a biomarker for the development of prostate neoplasia, and were not correlated with the clinicopathological characteristics of PCa

SmsDX suppresses the stimulative effect of MCM on the cell viability of LNCaP, DU145 and PC-3 cells.

<p>The effect of MCM and/or smsDX on cell viability was measured via CCK-8 assay. (A) LNCaP cells, (B) DU145 cells and (C) PC-3 cells were co-cultured with MCM and/or treated with smsDX for 24, 48 and 96 h. Co-culture with MCM significantly promoted the proliferation of PCa cells in a time-dependent manner, wheras smsdx could substantially attenuate this influence. (D) The effect of MCM and smsDX on colony formation. (E) The number of colonies, defined asclusters of at least 50 cells, was counted under microscope. The results represent means ± SD of three independent experiments. Data are presented as the means ± SD, *p<0.05, **p<0.01.</p

SmsDX inhibits the nuclear translocation of p65 induced by MCM in LNCaP, DU145 and PC-3 cells.

<p>(A, C, E)The expression of p65 in the cytoplasm was decreased after co-culture with MCM, smsDX attenuated this effect in LNCaP, DU145 and PC-3 cells. A-tubulin was chosen as the cytosolic protein loading control. (B, D, F) Co-culture with MCM increased the expression of nuclear p65, and treatment with smsDX weakened the effect of MCM. Histones were chosen as the nuclear protein loading control. The quantitation of the C-p65/ a-tubulin and N-p65/histone ratio was performed as densitometry analysis.</p

Simvastatin induced dose-dependent apoptosis in A498 and 786-O cells.

<p>(A) A498 and (B) 786-O cells were treated with simvastatin (0, 8 and 16 µM) for 48 hours and stained with FITC-annexinV and PI. The percentage of surviving cells was shown in the lower left quadrant; the percentage of early stage of apoptosis and late stage of apoptosis cells were shown in the lower right and upper right quadrants, respectively. (C, D) The quantification of apoptosis induced by simvastatin was calculated. Data is presented as mean ± SD of three independent experiments. * P<0.05, ** P<0.01.</p

Simvastatin depressed proliferation and metastasis of A498 cells that were transfected with AKT or ERK siRNA compared with the control cells (Mock).

<p>(A, B) A498 cells were transfected and treated with simvastatin (8 µM), and the levels of AKT and ERK were analyzed by western blotting with GAPDH as a control. After transfected with AKT or ERK siRNA, A498 cells were incubated in the absence or presence of simvastatin (8 µM) for 48 h. The cell viability was measured by MTT assay (C, D), and cell migration and invasion was measured by transwell assay (E, F). * p<0.05 or ** p<0.01, compared with the untreated cells (Mock). # p<0.05 or ## p<0.01, compared with the cells transfected with AKT or ERK siRNA.</p