Kindlin-2 promotes invasiveness of prostate cancer cells via NF-κB-dependent upregulation of matrix metalloproteinases.

Invasive progression is the major lethal cause of prostate cancer. In this study, we aimed to investigate the role of kindlin-2, an integrin-binding focal adhesion protein, in the regulation of invasiveness of prostate cancer. We found that downregulation of kindlin-2 using small interfering RNA (siRNA) technology significantly inhibited the invasion of PC-3 and DU-145 prostate cancer cells in a Matrigel Transwell assay. Conversely, overexpression of kindlin-2 promoted the invasiveness of prostate cancer cells. Kindlin-2 overexpression was found to activate nuclear factor (NF)-κB-dependent signaling and upregulate the expression of matrix metalloproteinase-9 (MMP-9) and MMP-2, whereas kindlin-2 silencing led to opposing effects on the expression of NF-κB and MMPs. Most importantly, kindlin-2-induced invasiveness was almost completely abolished by pretreatment with pyrrolidine dithiocarbamate (an inhibitor of NF-κB signaling) or co-transfection with MMP-9 or MMP-2 siRNA. Taken together, our data indicate that kindlin-2 promotes the invasiveness of prostate cancer cells largely through NF-κB-dependent upregulation of MMP-9 and MMP-2. Further studies are warranted to evaluate the significance of kindlin-2 as a therapeutic target for metastatic prostate cancer.

Percent free prostate-specific antigen is effective to predict prostate biopsy outcome in Chinese men with prostate-specific antigen between 10.1 and 20.0 ng ml(-1).

Percent free prostatic-specific antigen (%fPSA) has been introduced as a tool to avoid unnecessary biopsies in patients with a serum PSA level of 4.0-10.0 ng ml-1 , however, it remains controversial whether %fPSA is effective in PSA range of 10.1-20.0 ng ml-1 in both Chinese and Western population. In this study, the diagnostic performance of %fPSA and serum PSA in predicting prostate cancer (PCa) and high-grade PCa (HGPCa) was analyzed in a multi-center biopsy cohort of 5915 consecutive Chinese patients who underwent prostate biopsy in 22 hospitals across China from January 1, 2010 to December 31, 2013. The indication for biopsy was PSA>4.0 ng ml-1 or/and suspicious digital rectal examination. Total and free serum PSA determinations were performed by three types of electrochemiluminescence immunoassays with recalibration to the World Health Organization standards. The diagnostics accuracy of PSA, %fPSA and %fPSA in combination with PSA (%fPSA + PSA) was determined by the area under the receivers operating characteristic curve (AUC). %fPSA was more effective than PSA in men aged ≥60 years old. The AUC was 0.584 and 0.635 in men aged ≥60 years old with a PSA of 4.0-10.0 ng ml-1 and 10.1-20.0 ng ml-1 , respectively. The AUC of %fPSA was superior to that of PSA in predicting HGPCa in patients ≥60 years old in these two PSA range. Our results indicated that %fPSA is both statistically effective and clinical applicable to predict prostate biopsy outcome in Chinese patients aged ≥60 years old with a PSA of 4.0-10.0 ng ml-1 and 10.1-20.0 ng ml-1 .

[Effects of adenovirus-mediated PTEN on the proliferation of prostate cancer PC-3 cells and expressions of cyclin D1 and p21].

OBJECTIVE: To construct a recombinant adenovirus expression vector containing the anti-oncogene PTEN and to investigate the effects of the PTEN gene on the proliferation of prostate cancer PC-3 cells and the expressions of cyclin D1 and p21 in the PC-3 cells. METHODS: The PTEN gene was amplified from the rat hippocampus by RT-PCR and cloned into the shuttle plasmid pEN-TR2A. The plasmids were constructed and amplified in 293A cells. Prostate cancer PC-3 cells were cultured in vitro and infected with the adenoviral vector carrying the PTEN gene (Ad-PTEN). The up-regulation of the PTEN protein was measured by indirect immuno-fluorescence assay; the expressions of PTEN, cyclin D1 and p21 in the cells infected with Ad-PTEN and Ad-LacZ were determined by RESULTS: The Western blot; and the effect of PTEN on the cell proliferation was detected by MTT assay and plate colony formation. recombinant adenoviral vector Ad-PTEN was successfully constructed. Western blot showed a significantly increased expression of the PTEN protein in the PC-3 cells infected with Ad-PTIEN (0.215 +/-0.065) as compared with that in the control ([0.052 +/-0.009], t = 4. 30, P <0.05) and the Ad-LacZ group ( [0. 056 +/- 0.008 ] , t =4.21, P <0.05). The expression of cyclin D1 was significantly lower in the Ad-PTEN-infected PC-3 cells (0. 256 +/- 0. 072) than in the control ( [0. 502 +/- 0. 087 ], t = 3.77, P < 0.05) and the Ad-LacZ group ([0.498 +/-0.081] , t =3.87, P <0.05), while the expression of p21 remarkably higher in the Ad-PTEN-infected PC-3 cells (0.589 +/-0. 076) than in the control ([0. 146 +/-0.026] , t = 9.55, P<0. 01) and the Ad-LacZ group ([0. 163 +/-0. 024] , t = 9.26, P <0.01). Ad-PTEN significantly inhibited the growth of the PC-3 cells (21.98%) at 48 h (t = 6.80, P <0.01). The colony formation rate of the PC-3 cells was (37.4 +/-4. 18)% in the Ad-PTEN group, significantly lower than (54.9 +/-4.81)% in the control (t =4.76, P<0.01) and (56.5 +/- 5.42)% in the Ad-LacZ group (t=4.83, P<0.01). CONCLUSION: The expression of PTEN induced by Ad-PTEN can significantly inhibit the proliferation of PC-3 cells, down-regulate the expression of cyclin D1, and up-regulate the expression of p21.

Role of PSA-related variables in improving positive ratio of biopsy of prostate cancer within serum PSA gray zone.

OBJECTIVE: The aim of this study was to assess the validity of predictors of prostate biopsy outcome in order to improve their positive predictive value (PPV). The related variables of PSA such as free/total PSA ratio (F/T), complexed/total PSA ratio (C/T), density of PSA (PSAD) were studied to improve early diagnosis for early PCa within serum PSA gray zone. METHODS: Samples of venous blood were collected early in the morning from 459 patients, aged 50 to 88 years (average 70.5 years old), whose serum T-PSA was between 4 to 10 ng/mL (153 cases of Pca, 306 cases of BPH). Serum T-PSA, F-PSA and C-PSA were determined by ELISA method; prostate volume was measured by transrectal ultrasound, then prostate biopsy was taken for all patients. RESULTS: Out of f 459 patients, 153 cases of Pca and 306 of BPH were diagnosed by prostate biopsy, whose mean values of T-PSA were 7.2 ± 1.25 ng/mL and 6.8 ± 0.96 ng/mL, respectively (P<0.05), of F/T were 0.117 ± 0.018 and 0.196 ± 0.021 (P<0.01), and of C/T they were 0.890 ± 0.023 and 0.754 ± 0.015 (P<0.01), of PSAD were 0.197 ± 0.023 and 0.106 ± 0.019 (P<0.01). The threshold of F/T>0.16 diagnosed 91 cases of cancer (59.5%), of C/T>0.84 88 cases (57.5%), of PSAD>0.15 94 cases (61.4%), and especially, of F/T>0.16 combined PSAD>0.15 80 cases whose sensitivity, specificity, accuracy, positive predictive values were 52.3%, 76.1%, 68.2%, 52.3%, respectively (P<0.05). CONCLUSION: There are some defects only in taking T-PSA as a biomarker for the diagnosis of PCa within serum PSA gray zone, and the threshold of F/T>0.16 combined PSAD>0.15 may be a new and significant indicator to provide a better basis for the diagnosis of early PCa.

Up-regulation of pVHL along with down-regulation of HIF-1α by NDRG2 expression attenuates proliferation and invasion in renal cancer cells.

The majority of renal cell carcinomas (RCCs) are characterized by loss of function of the tumor suppressor gene von Hippel Lindau (VHL), which acts as ubiquitin ligase for hypoxia-inducible factor-1α (HIF-1α). In the absence of VHL, HIF-1α protein becomes stabilized and contributes to tumorigenesis. Recent data demonstrate the antitumor efficacy of VHL promoter in RCC cells. This study demonstrates that N-Myc downstream-regulated gene 2 (NDRG2) is a potential regulator of VHL. NDRG2 is involved in proliferation and invasion of cancer cell, furthermore it is frequently down-regulated in renal cell carcinoma. Herein we evaluated the effect of NDRG2 overexpression on proliferation and invasion in human renal cancer cells. The human renal cancer cell line 786-O and A498 were infected with Ad-NDRG2 or Ad-LacZ. Overexpression of NDRG2 not only inhibited the growth of the cells, but also suppressed the invasion. Further study showed that the tumor suppressor gene VHL were up-regulated, whereas transcription factor HIF-1a and vascular endothelial growth factor (VEGF) were down-regulated in 786-O cells infected by Ad-NDRG2. Finally, in a nude mouse model, intratumoral injections of Ad-NDRG2 every 3 days for a total of seven times significantly inhibited the growth and angiogenesis of xenografted 786-O tumors. In conclusion, these data indicate that NDRG2 may be involved in proliferation and invasion by impacting the expression of VHL and HIF-1α. NDRG2 may be an attractive therapeutic target for renal cell carcinoma.

Flank suspended supine position for percutaneous nephrolithotomy.

AIMS: Prone and supine positions for percutaneous nephrolithotomy are widely used but have their drawbacks. We report a new positioning method called "flank suspended supine position" (FSSP) for PCNL and describe our experience with PCNL in this position to evaluate its safety and efficacy. METHODS: Retrospective study of 150 cases of renal stone patients treated with PCNL in a new position called flank suspended supine position (FSSP) from June 2009 to July 2010. All patients were treated with PCNL in FSSP under epidural anesthesia. Operation time, bleeding rate, stone free rate, and complications were recorded. RESULTS: All patients tolerated FSSP. Mean operation time was 78.29±26.13 min. Initial stone-free rate was 83%. For those with residual stones (26 cases), 18 were stone-free after a second PCNL, 8 after extracorporeal shock wave lithotripsy (ESWL). Mean hospital stay was 7.63±2.39 days. No penetrating injury of the pleural cavity or injury to visceral organs was reported. SUMMARY: FSSP is an effective and safe position for PCNL in our hands and its effectiveness relative to traditional prone position needs to be determined in future randomized studies.

[Effects of staurosporine on the proliferation and apoptosis of prostate cancer PC-3 cells].

OBJECTIVE: To investigate the effects of staurosporine (ST) on the proliferation and apoptosis of prostate cancer PC-3 cells. METHODS: Prostate cancer PC-3 cells were treated in vitro with ST at 10(-8) mol/L. The expressions of cyclin A and cyclin D1 proteins in the cells were detected by Western blot, the effect of ST on the proliferation of the cells determined by MTT assay and plate colony formation, the apoptosis of the cells examined by flow cytometry, and their morphological changes observed under the light microscope. RESULTS: ST treatment markedly decreased the expressions of cyclin A and cyclin D1 in the PC-3 cells, and significantly inhibited the growth of the PC-3 cells (19.35%) at 48 h. (F = 31.06, P < 0.01). The colony formation rate of the PC-3 cells was (37.10 +/- 3.43) % in the ST group, significantly lower than (64.80 +/- 4.34) % in the control (chi2 = 14.59, P < 0.05) and (62.80 +/- 4.36) % in the DMSO group (chi2 = 12.50, P < 0.05), while the apoptosis rate of the cells was remarkably higher in the ST group ([19.6 +/- 2.20] %) than in the control ([5.33 +/- 1.40] %) and the DMSO group ([5.50 +/- 0.96] %) (F = 104.36, P < 0.01). Under the light microscope, the ST-treated cells were round with indistinct margins as compared with those of the other two groups. CONCLUSION: ST could significantly inhibit the proliferation and induce the apoptosis of PC-3 cells.

[Effects of intraprostatic injection of botulinum toxin A (BTX-A) on benign prostate hyperplasia].

OBJECTIVE: To investigate the effects of intraprostatic injection of botulinum toxin A (BTX-A) on benign prostate hyperplasia (BPH) in rats. METHODS: Models of BPH were established in adult male Sprague-Dawley rats by injection of testosterone propionate, and then divided into three BTX-A groups, injected with BTX-A into the ventral prostate at the doses of 5 U, 10 U and 20 U, a negative control group, injected with saline only, and a sham operation group, with 12 in each. The prostates of the animals were harvested at 2 or 4 weeks after the injection, their volumes and weights measured, histological changes examined by HE staining, and glandular and interstitial areas semi-quantified by the image analysis system. RESULTS: Two rats died in the 20 U group within 3 days after BTX-A injection. Compared with the saline group, the 5 U, 10 U and 20 U BTX-A groups showed significant decreases in prostatic volume (P < 0.01, 0.01 and 0.05), weight, and glandular and interstitial areas as well as atrophic epithelia in the glandular tube at 2 weeks. These changes were lessened at 4 weeks, especially in the 5 U group. CONCLUSION: Intraprostatic injection of BTX-A induces obvious atrophy and histological changes of the prostate, but meanwhile may potentially result in death at a large dose.

[The mRNA expression of P16(ink4a) and HST2 in benign prostatic hyperplasia tissues: a pilot study].

OBJECTIVE: To investigate the expressions of the aging gene P16(ink4a) and anti-aging gene HST2 in benign prostatic hyperplasia (BPH). METHODS: Twenty-three BPH and eighteen normal prostate specimens were collected and total RNA was extracted, followed by the reverse transcriptase polymerase chain reaction (RT-PCR). The expressions of P16(ink4a) was detected by semi-quantitative analysis in BPH and normal prostate tissues. RESULTS: P16(ink4a) mRNA, rather than HST2, was expressed in the BPH and normal prostate tissues. Semi-quantitative analysis showed that the P16(ink4a) mRNA expression in the normal prostate tissues (0.4868 +/- 0.545 was significantly higher than in the BPH tissues (0.2783 +/- 0.0268, with a statistical difference in between (P < 0. 05). CONCLUSION: P16(ink4a) might play an important role in the pathogenesis of BPH and is probably one of the factors of cell aging escape.

Heparanase promotes bone destruction and invasiveness in prostate cancer.

Heparanase is an endoglycosidase that plays an important role in angiogenesis and metastasis of cancer. Herein we evaluate the effect of heparanase overexpression on invasiveness and bone destruction in prostate cancer bone metastases. The human prostate cancer cell line PC-3 was stably transfected with a plasmid containing the cDNA for human heparanase or with the vector alone as a control. Overexpression of heparanase did not affect the growth of PC-3 cells, but did promote invasiveness of the cells in an in vitro assay. Both cell types were injected into the tibias of nude mice. Four weeks later, the mice were examined radiologically prior to sacrifice and samples of leg tissue were taken to investigate bone destruction and metastasis. Mice injected with PC-3 cells overexpressing heparanase had more severe bone destruction and larger, more invasive, tumors. These results demonstrate that heparanase overexpression can facilitate tumor invasion and accelerate bone destruction caused by prostate cancer bone metastasis.

Histone deacetylase inhibitor trichostatin A inhibits the growth of bladder cancer cells through induction of p21WAF1 and G1 cell cycle arrest.

PURPOSE: To investigate whether Trichostatin A (TSA) possesses antitumor activity against human bladder cancer cells, and if any, its mechanism. MATERIALS AND METHODS: A human bladder cancer cell line, BIU-87, was treated with different concentrations of TSA. After treatment, cell growth was measured by MTT assay. Cell apoptosis and cell cycle changes were examined by means of flow cytometry (FCM). Apoptosis was confirmed by apoptotic ladder formation assay. mRNA expression of p21WAF1 and p53 was assessed by differential reverse transcription-polymerase chain reaction. RESULTS: Trichostatin A significantly inhibited the proliferation of bladder cancer cell at nanomolar concentrations in a time- and dose-dependent fashion. TSA treatment caused cell cycle arrest at the G1 phase and increased apoptotic cell death as shown by FCM and DNA fragmentation analysis, accompanied by increased p21WAF1 mRNA expression. In addition, TSA treatment did not alter p53 mRNA expression. CONCLUSION: Our results indicate that TSA is able to inhibit bladder cancer cell growth in vitro, possibly through p21WAF1 mediated cell cycle arrest and apoptotic cell death. This study suggests that TSA may be a potential therapeutic agent for the treatment of bladder cancer.

Renal pedicle lymphatic disconnection for chyluria via retroperitoneoscopy and open surgery: report of 53 cases with followup.

PURPOSE: We present our experience with retroperitoneoscopic renal pedicle lymphatic disconnection. We compared the clinical efficacy of this treatment for chyluria with that of open surgery. MATERIALS AND METHODS: From January 1998 to June 2004, 53 patients (55 renal units) with chyluria underwent renal pedicle lymphatic disconnection via the retroperitoneoscopic and conventional open approaches. The diagnosis of chyluria was confirmed by the ether test and the side of chylous reflux was determined by cystoscopy. Operative time, intraoperative blood loss, postoperative intestinal recovery and hospital stay were evaluated. Increases in hemoglobin and serum albumin were compared before and after surgery during followup. RESULTS: Retroperitoneoscopic renal pedicle lymphatic disconnection or open surgery was performed successfully in all patients. In terms of operative time, intraoperative blood loss, postoperative intestinal recovery and hospital stay retroperitoneoscopy was superior to conventional open surgery. During retroperitoneoscopy the inferior vena cava was injured in 1 case but repaired successfully by laparoscopy without conversion to open surgery. Postoperative gross hematuria in 1 case disappeared 4 days later. In the open surgery group the renal segmental artery was inadvertently injured in 1 case and anastomosis was performed successfully. Wound healing was delayed in 1 case due to hypoalbuminemia. Recurrence developed in 2 patients during the 6 to 84-month followup. CONCLUSIONS: Retroperitoneoscopic renal pedicle lymphatic disconnection for chyluria has the advantages of minimal invasion and rapid recovery compared with open surgery.

[The changes of IGF-I in testis and epididymis on a rat model with oligozoospermia/azoospermia induced by cyclophosphamide].

OBJECTIVE: To evaluate the effect of the levels of IGF-I in the epididymis and the expression of IGF-I in the testis of adult male rat after the administration of cyclophosphamide. METHODS: Ninety-six male adult rats (8 weeks age) were divided into 6 groups. The doses given to the rats of the groups 1 to 5 were 10, 20, 40, 80 and 100 mg/(kg x d), respectively. The remaining group was served as control. All those rats were sacrificed and IGF-I were quantitatively determined by ELISA techniques 2 and 4 weeks after the administration of the drug (by gastric fudge). Immunohistochemical SP technique was used to examine expression of IGF-I in rat testis. RESULTS: The levels of cell factors (IGF-I) in the epididymis of the rats were gradually reduced with the increasing time and dose after administration of the drug. In the mean time the expression of IGF-I in the tissues of the testis of those rats were also gradually reduced. CONCLUSION: In the time of oligozoospermia/azoospermia induced by the administration of cyclophosphamide, the expression levels of IGF-I in the genetic system were significantly reduced. The possible mechanism of these changes could be attributed to the lower spermatogenesis function of the testis caused by the administration of cyclophosphamide.

Comparison of retroperitoneoscopic nephrectomy versus open approaches to nonfunctioning tuberculous kidneys: a report of 44 cases.

PURPOSE: We retrospectively investigated the advantages of retroperitoneoscopic nephrectomy for nonfunctioning tuberculous kidneys by comparing its clinical results, operative methods and skills with those of open nephrectomy. MATERIALS AND METHODS: Clinical data on 22 patients with nonfunctioning tuberculous kidneys who underwent retroperitoneoscopic nephrectomy, including simple and subcapsular nephrectomy, were compared with those on 22 who underwent open nephrectomy for a similar indication during the same period. Results in the 2 groups were analyzed. RESULTS: There was no statistical difference between the retroperitoneoscopy and open surgery groups with regard to patient age, sex or mean operative time +/- SD (93.0 +/- 12.6 vs 92.6 +/- 35.5 minutes). Mean blood loss was significantly less in the retroperitoneoscopy group than in the open surgery group (78.3 +/- 60.6 vs 160 +/- 120.0 ml). Mean hospital stay after operation was notably shorter in the retroperitoneoscopy group compared with the open surgery group (3.3 +/- 0.9 vs 9.1 +/- 0.8 days). The mean analgesic requirement for opioids and diclofenac sodium was also lower in the retroperitoneoscopy group than in the open surgery group (0 vs 2.1 +/- 0.9 and 5.2 +/- 1.1 vs 5.8 +/- 1.3 doses, respectively). CONCLUSIONS: Retroperitoneoscopic nephrectomy for renal tuberculosis has several advantages over open nephrectomy, namely a smaller wound, less blood loss and more rapid recovery. It may provide a safe and reliable method for treating refractory renal tuberculosis clinically.