Knockdown of ZFX suppresses renal carcinoma cell growth and induces apoptosis.

The ZFX (zinc finger protein, X-linked) gene located on the human X chromosome controls the self-renewal of embryonic and hematopoietic stem cells as a transcriptional regulator. Recently, studies have affirmed that ZFX is associated with several human cancers, including lymphoma, laryngeal squamous cell carcinoma, prostate cancer, and liver cancer, which suggests ZFX as a potential therapeutic target in cancer. However, the functional role of ZFX in human renal cancer remains unclear. Herein, we detected the expression of ZFX in 42 patients with renal cancer and found the expression of ZFX was specifically upregulated in cancer tissues at the mRNA and protein levels. Moreover, we employed lentivirus-mediated short hairpin RNA (shRNA) to knock down ZFX expression in two human renal cell carcinoma cell lines, 786-0 and ACHN. Functional analysis indicated that ZFX silencing significantly inhibited renal cell carcinoma cell proliferation and cell cycle progression, probably because of suppression of CDK4 and cyclin D1, and induced apoptosis via activation of Bax, Caspase 3, and PUMA in a p53-dependent manner. Our findings suggest that knockdown of ZFX by shRNA may be a potential therapeutic approach for the treatment of renal cancer.

Selective transurethral resection of the prostate combined with transurethral incision of the bladder neck for bladder outlet obstruction in patients with small volume benign prostate hyperplasia (BPH): a prospective randomized study.

PURPOSE: Transurethral resection of the prostate (TURP) has a high failure rate in patients with small volume benign prostate hyperplasia (BPH) with bladder outlet obstruction (BOO). We describe and report the results of an alternative surgical method, selective transurethral resection of the prostate (STURP) in combination with transurethral incision of the bladder neck (TUIBN). METHODS: Patients were randomized to receive TURP or STRUP+TUIBN in combination with TUIBN. Maximum urinary flow rate (Qmax), voided volume, and post voiding residual volume (PVR) were assessed at baseline and at 1, 3, and 6 months after surgery. Efficacy of treatment was assessed by lower urinary tract symptoms and IPSS. RESULTS: Sixty three patients received STRUP+TUIBN and 61 received TURP. Surgical time, amount of prostate tissue resected, and blood loss was the same in both groups (all, p>0.05). The mean duration of follow-up was 9.02 and 8.53 months in patients receiving TURP and STRUP+TUIBN, respectively. At 6 months postoperatively, IPSS was 4.26±1.22 and 4.18±1.47 in patients receiving TURP and STRUP+TUIBN, respectively (p>0.05), and the Qmax in patients receiving STRUP+TUIBN was markedly higher than in those receiving TURP (28.28±6.46 mL/s vs. 21.59±7.14 mL/s; p<0.05). Bladder neck contracture and urinary tract infections were observed in 3 and 5 patients receiving TURP, respectively, and none in STURP. CONCLUSIONS: STRUP+TUIBN may offer a more effective and safer alternative to TURP for small volume BPH patients.

Association of BLCA-4 hypomethylation in blood leukocyte DNA and the risk of bladder cancer in a Chinese population.

Global DNA hypomethylation has been associated with increased risk for cancers of the colorectum, bladder, breast, head and neck, and testicular germ cells. The aim of this study was to examine whether global hypomethylation measured at BLCA-4 repeat regions through bisulfite pyrosequencing in blood leukocyte DNA is associated with the risk of bladder cancer(BC). A total of 312 bladder cancer patients and 361 healthy control subjects were included in Chongqing, China. Global methylation in blood leukocyte DNA was estimated by analyzing BLCA-4 repeats using bisulfite-polymerase chain reaction (PCR) and pyrosequencing. The median methylation level in BC cases (percentage of 5-methylcytosine (5 mC) = 75.7 %) was significantly lower than that in controls (79.7 % 5 mC) (P = 0.002, Wilcoxon rank-sum test). The odds ratios (ORs) of BC for individuals in the third, second, and first (lowest) quartiles of BLCA-4 methylation were 1.2 (95 % confidence interval (CI) 0.8-1.9), 1.6 (95 % CI 1.1-2.3), and 2.7 (95 % CI 1.5-3.8) (P for trend <0.001), respectively, compared to individuals in the fourth (highest) quartile. A 2.1-fold (95 % CI 1.5-2.8) increased risk of BC was observed among individuals with BLCA-4 methylation below the median compared to individuals with higher (>median) BLCA-4 methylation. Our results demonstrate for the first time that individuals with global hypomethylation measured in BLCA-4 repeats in blood leukocyte DNA have an increased risk for BC. Our data provide the evidence that BLCA-4 hypomethylation may be a useful biomarker for poor prognosis of patients with BC.

Suppression of CX43 expression by miR-20a in the progression of human prostate cancer.

The aberrant expression of microRNAs (miRNAs) has been found in various types of cancer. The present study found miR-20a to be significantly upregulated in prostate cancer compared with normal prostate tissues. The proliferation and colony formation assays revealed that the downregulation of miR-20a by miR-20a inhibitor suppresses the proliferation of MDA-PCa-2b cells in vitro and also inhibits tumor growth in vivo. Furthermore, a gap junction protein, α 1 (CX43), was identified as a direct target gene of miR-20a. The upregulation of CX43 was detected in MDA-PCa-2b cells after treatment with miR-20a inhibitor both in vitro and in vivo. In conclusion, the findings show that miR-20a significantly contributes to the progression of prostate cancer by targeting CX43.

Resection of a giant renal angiomyolipoma in a solitary kidney with preoperative arterial embolization.

Renal angiomyolipoma is a type of benign tumor that occurs sporadically in addition to being associated with tuberous sclerosis. Preoperative embolization of large tumors is important to avoid excessive blood loss during surgery. We reported a patient with a 5505-g giant renal angiomyolipoma in a solitary kidney. The patient was treated with preoperative embolization and radical nephrectomy without complications. This type of treatment for an enormous angiomyolipoma can reduce the risk of uncontrolled hemorrhage caused by rupture of the tumor during the operation and should be considered for the treatment of similar tumors.

[Study on the neurophysiologic of detrusor overactivity due to partial bladder outflow obstruction].

OBJECTIVE: To study the neurophysiologic of detrusor overactivity (DO) due to partial bladder outflow obstruction (PBOO). METHODS: Twenty four female Wistar rats with DO caused by PBOO were studied simultaneously with ten sham-operated rats. An electrophysiological multi-channel simultaneous recording system was used to record pelvic afferent fiber potentials as well as the pudendal nerve motor branch potentials, external urethral sphincter electromyogram (EUS EMG) and abdominal muscle EMG during filling cystometry. To test the effect of the unstable contraction in DO rats after the decentralization of the central nervous system, DO rats were studied the changes of the unstable contraction after transection of the spinal cord (T(8) level), pelvic nerve, the sympathetic trunk, and the pudendal nerve. RESULTS: The incidence of DO was 62.5% in filling cystometry. During filling cystometry, there are two type of DO contraction according to the changes of pelvic afferent fiber signals, the relevant nerves and muscles responses: the small pressure of the unstable contraction (S-DO) and the big pressure of the unstable contraction (B-DO). For the B-DO, there were significant changes in the recordings of pelvic afferent fiber, the motor branch of the pudendal nerve, EUS EMG, and abdominal muscle EMG. While all these differences have not been recorded during S-DO. Both the filling-voiding cycle and the unstable contraction of B-DO were eliminated and the base line of bladder pressure increased after T(8) spinal cord transection. While the S-DO was not affected by such transection. When bladder relevant nerves were transected by the sequence of the pelvic nerve, the sympathetic trunk, and the pudendal nerve, the filling-voiding cycle was eliminated. The base line of bladder pressure increased significantly. No B-DO was recorded, but the S-DO still existed. CONCLUSION: There are some bladder-genic factors take part in the DO contractions induced by PBOO.

[Correlation between activation of L5-S2 spinal cord astrocytes and effect of substance P in chronic prostatitis pain].

OBJECTIVE: To observe the expressions of the substance P (SP) mRNA and neurokinin-1 receptor (NK-1R) in the posterior horn of the L5 - S2 spinal cord in the rat model of chronic prostatitis pain, and to investigate the changes in the activation of astrocytes and influence of SP on this activation in rat spinal cord astrocytes cultured in vitro. METHODS: The rat model of chronic prostatitis pain was established by injection of complete Freund's adjuvant (CFA) and assessed by the tail flick threshold test, the control rats injected with sodium chloride and all observed at 0, 14 and 28 days. Changes in the expressions of SP mRNA, NK-1R, glial fibrillary acidic protein (GFAP), tumor necrosis factor-alpha (TNF-alpha) and inducible nitric oxide synthase (iNOS) in the posterior horn of the L5 - S2 spinal cord were detected by RT-PCR and Western blot. Rat spinal cord astrocytes were cultured in vitro and divided into a control group, cultured with ITS cell culture fluid, and two experiment groups, with Group 1 stimulated with SP at the concentration of 10(-9) - 10(-6) mol/L for 12 hours followed by determination of the expressions of TNF-alpha, IL-1beta, NO and NOS by ELISA and nitrate reductase and colorimetric methods, and Group 2 at 10(-7) mol/L for 0, 24, 48 and 72 hours followed by detection of the GFAP expression by Western blot. RESULTS: The expressions of SP mRNA, NK-1 R, GFAP, TNF-alpha and iNOS in the posterior horn of the L5 - S2 spinal cord were obviously higher in the rat prostatitis pain models than in the controls, successively higher at 28 than at 14 and 0 d (P < 0.01), and so was the expression of GFAP at 28 than at 14 d in the experiment groups (P < 0.05). SP induced a gradual increase at 10(-7) mol/L in the expression of GFAP in the spinal cord astrocytes at 0 -72 h, significantly different from that of the control group (P < 0.01), and it promoted the excretion of TNF-alpha and IL-1beta and the activity of NO and NOS at 10(-9) - 10(-6) mol/L at 12 h in a concentration-dependent manner, with marked differences between the experiment and control groups (P < 0.01, P < 0.05). But a decreased excretion of IL-1 beta was observed in the 10(-6) mol/L group, though with no significant difference from the control (P > 0.05). CONCLUSION: Chronic prostatitis pain could upregulate the expressions of the excitatory transmitter SP and receptor in the L5 - S2 spinal cord, and result in the activation of astrocytes and increased excretion of proinflammatory cytokines, which may be associated with the persistence and generalization of prostatitis pain.

[Morphological study on the role of ICC-like cells in detrusor neuro-modulation of rat urinary bladder].

OBJECTIVE: To investigate the role of ICC-like cells in bladder neuromodulation in rat urinary bladder. METHODS: 14 SD rats and 1 guinea pig were sacrificed in this study. The ultra structural relationships among interstitial cells, nerves and detrusor smooth muscle cells (DSMCs) of urinary bladder were investigated by transmission electron microscopy (TEM). c-kit immunofluorescence was used to identify ICC-like cells in SD rat urinary bladder and the structural relationship between ICC-like cells and nerve terminals was studied by immunofluorescence (double-label). RESULTS: Gap junction between ICC-like cells and DSMCs was confirmed by TEM. ICC-like cells were very close apposition with nerve terminals under TEM. ICC-like cells were identified to exist in sub-urothelium layer, along the longitude of smooth muscle bundles and among detrusor smooth muscle in SD rat urinary bladder by c-kit immunofluorescence. Double-labeled tissue with c-kit and PGP9.5 antibodies also showed that ICC-like cells were very close apposition with nerve terminals in SD rat bladder. CONCLUSIONS: Morphological study indicated that ICC-like cells in rat urinary bladder may play an important role in detrusor neuromodulation. Further study on function will be helpful for elucidating the mechanism of bladder neuromodulation clearly.