[Show abstract][Hide abstract] ABSTRACT: MicroRNAs (miRNAs) are small non-coding RNAs that silence their cognate target genes by specifically binding and cleaving messenger RNAs or inhibiting their translation. In this study, we explored whether miRNA-141 and miRNA-200b are involved in regulation of the invasive ability and epithelial-mesenchymal transition (EMT) of bladder cancer cells in vitro. We also evaluated their potential as biomarkers for deciding the extent of pelvic lymph node dissection (PLND) required during radical cystectomy.
Pri- and anti-miR cell lines were constructed. The invasive capacity of the cells was tested using a cell invasion assay. The MMP-2, MMP-9 and EMT-related markers were validated through Western blotting analysis. Seventy-eight urine samples from patients undergoing cystectomy and super-extended lymph node dissection were evaluated by qRT-PCR.
Loss of expression of miRNA-141 and miRNA-200b was associated with increased invasion and migration ability, upregulated MMP-2, MMP-9, vimentin and N-cadherin expression, and downregulated E-cadherin expression in bladder cancer cell lines. Urine miRNA-141 and miRNA-200b levels could discriminate patients with lymph node metastasis from those who were lymph node negative (AUC: 0.704 and 0.674, respectively).
MiRNA-141 and miRNA-200b play important roles in the invasive ability and EMT phenotype of bladder cancer. Detection of miRNA-141 and miRNA-200b can help to identify patients undergoing cystectomy who are likely to have lymph node metastasis, and therefore those who may benefit from super-extended PLND.
BMC Cancer 12/2015; 15(1):1110. DOI:10.1186/s12885-015-1110-7 · 3.32 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Afatinib is a highly selective, irreversible inhibitor of the epidermal growth factor receptor (EGFR) and human EGFR 2 (HER-2). Although preclinical and clinical studies have indicated that afatinib has antitumor activity and clinical efficacy in non-small cell lung carcinoma, head and neck squamous cell carcinoma and breast cancer, there are few studies investigating its inhibitory effect on human bladder carcinoma cells. In this study, the antitumor effect of afatinib was investigated on the T24 bladder cancer cell line. The T24 bladder cancer cell line was treated with afatinib at various concentrations (0, 1, 5, 10 and 20 µmol/l). MTT assay was used to estimate the proliferation of the T24 cells; flow cytometric analysis was used to estimate the effect of afatinib on T24 cell apoptosis; cell invasion ability was assessed by a Transwell invasion assay; and western blot analysis was used to detect the expression of Bcl-2, Bax, Akt, extracellular-signal-regulated kinase (ERK)1/2, matrix metalloproteinase (MMP)-2 and MMP-9. The MTT assay demonstrated that afatinib inhibited the proliferation of T24 cells in a dose- and time-dependent manner. Flow cytometric analysis revealed that the cell apoptosis rate increased as the concentration of afatinib increased. The cell invasion assay indicated that afatinib treatment significantly inhibited the invasive behavior of T24 cells in a dose-dependent manner. Western blot analysis showed that with increasing afatinib concentrations, Bcl-2, phosphorylated (p)-ERK1/2, p-Akt, MMP-2 and MMP-9 expression levels were significantly decreased, whereas total (t)-ERK1/2 and t-Akt expression levels remained basically unchanged, and Bax expression levels were greatly increased. The results indicate that afatinib inhibits the proliferation and invasion of T24 cells in vitro and induces the apoptosis of these cells by inhibiting the EGFR signaling network.
Experimental and therapeutic medicine 02/2015; 9(5). DOI:10.3892/etm.2015.2314 · 0.94 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: To evaluate the efficacy and safety of self-retaining barbed suture (SRBS) application during laparoscopic partial nephrectomy (LPN), by assessing perioperative outcomes.
Data from consecutive patients who underwent retroperitoneal LPN from January 2008 to December 2013 were retrospectively collected. Patients were divided into two groups according to suture techniques for renorrhaphy: the sliding clip technique and SRBS. The SRBS cases (Group 2 [G2]) were 1:1 matched with cases in the sliding clip group (Group 1 [G1]) for the PADUA score. Patient characteristics, perioperative outcomes, and renal function changes were compared between the groups.
In total, 41 patients in G1 and 41 patients in G2 successfully underwent LPN. Patient characteristics, operative time, and complication rate were similar for both groups. Mean warm ischemia time was significantly shorter for the SRBS group (G1 versus G2, 27.5 versus 20.7 minutes; P<.05). The estimated blood loss was similar in both groups. An improved early affected renal function recovery was observed in the SRBS group (percentage of glomerular filtration rate reduction for G1 versus G2, 29% versus 20.8%; P<.05).
The SRBS application offers an effective and safe renal parenchyma repair. In addition, SRBS appears to significantly minimize the warm ischemia injury and results in superior short-term renal function preservation.
[Show abstract][Hide abstract] ABSTRACT: Fast-track surgery (FTS), which combines various techniques with evidence-based adjustments, is aimed to reduce postoperative morbidity, attenuate surgical stress response, thereby accelerating recovery and shorting length of stay. To further investigate the effectiveness of fast-track surgery, we compared the short-term outcomes of laparoscopic radical cystectomy and ileal conduit diversion for Chinese bladder cancer patients with FTS or with CS in our hospital. Patients with bladder cancer were included and divided into two consecutive groups: CS group and FTS group. Duration to first flatus and regular diet, postoperative hospital days, hospital expense, incidence of complications and postoperative surgical stress response were compared. There was no significant difference between the two groups in age, sex, BMI and postoperative TNM classification. Compared with the CS group, the FTS group had significantly shorter duration to first flatus, time to regular diet, postoperative hospital days and hospital expense, less complications, lower white blood count (WBC) and serum of C-reactive protein (CRP) on postoperative day 5 and 7. Our study indicates that FTS program is safe and efficacious for Chinese patients undergoing laparoscopic radical cystectomy and ileal conduit diversion. It can accelerate recovery, reduce stress action, shorten postoperative hospitals days and reduce hospital expenses.
[Show abstract][Hide abstract] ABSTRACT: Wild-type p53-induced phosphatase (Wip1 or PPM1D) has been reported to be aberrantly expressed in various cancers and correlated with the malignant behavior of cancer cells. However, the function of Wip1 in RCC remains unclear. The present study investigated its abnormal expression and dysfunctions in clear cell renal cell carcinoma (ccRCC) in vitro. With the combination of immunohistochemistry, western blotting, immunofluorescence, qRT-PCR, and cell proliferation, migration and invasion assays, we found that levels of Wip1 mRNA and protein were dramatically increased in human ccRCC tissues (P<0.001 for both), and upregulation of Wip1 was significantly associated with depth of invasion (P<0.001), Distant metastasis (P = 0.001), lymph node status (P<0.001) and Fuhrman grade (P<0.001). Wip1 knockdown inhibited the proliferation, migration and invasion of 786-O and RLC-310 cells, whereas Wip1 overexpression promoted the growth and aggressive phenotype of 786-O and RLC-310 cells in vitro. The uni- and multivariate analyses indicated that expression of Wip1 was an independent predictor for survival of ccRCC patients (P = 0.003, P = 0.027 respectively). Wip1- negative patients had a higher tumor-free/overall survival rate than patients with high Wip1 expression (P = 0.001, P = 0.002 respectively). Overexpression of Wip1 is useful in the prediction of survival in ccRCC patients.
PLoS ONE 10/2014; 9(10):e110218. DOI:10.1371/journal.pone.0110218 · 3.23 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Objectives
We seek to summarize the clinicopathologic characteristics and prognostic factors of sarcomatoid renal cell carcinoma (SRCC), an uncommon type of renal cell carcinoma.
Methods and materials
Between 2004 and 2012, 23 patients with SRCC were treated at a large urology center in south central China. We collect patient’s clinicopathologic features from medical records to assess diagnosis, prognostic factors and efficacy of systemic therapy. Clinical data were absent in 3 cases, and 20 patients were enrolled in the final study.
Immunohistochemically, almost all SRCC expressed cytokeratin (91 %), epithelial membrane antigen (87 %) and vimentin (100 %). Sarcomatoid differentiation occurs in various kinds of subtypes of RCC with almost the same probability. The median tumor size was 10.5 cm. The CT findings of these tumors revealed low-density (n = 5; 25 %) or mixed (n = 15; 75 %) masses with necrotic areas and often showed an infiltrative morphology (n = 15; 75 %). All 20 cases demonstrated heterogeneous enhancement, and eleven (55 %) cases demonstrated >50 % necrosis. Six cases complicated with calculus and hydronephrosis. Sixteen (80 %) patients demonstrated invasions of tissues localized in Gerota’s fascia, and 8 (40 %) tumors invaded beyond Gerota’s fascia. Fifteen (75 %) patients demonstrated lymph node metastasis, and sixteen (80 %) patients had distant metastasis. Five patients received systemic therapy, and one patient given high-dose interferon-α had a completely response, and one patient received chemotherapy based on gemcitabine had partial response. The median overall survival of all patients was 5.8 months. Patients without distant metastasis had a median overall survival of 35 months compared with 3 months of those with distant metastasis (P
Journal of Cancer Research and Clinical Oncology 09/2014; 141(2). DOI:10.1007/s00432-014-1740-1 · 3.01 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Transforming growth factor‑β1 (TGF‑β1) is involved in the migration and metastases of bladder cancer. The present study was designed to investigate whether TGF‑β1 is able to induce epithelial‑mesenchymal transition (EMT) and the upregulation of matrix metalloproteinase‑16 (MMP‑16), and to identify an association between EMT and MMP‑16 in bladder cancer. Following TGF‑β1 treatment, samples of HTB9 and T24 bladder cancer cells were collected at various time points. Western blotting and quantitative polymerase chain reaction (qPCR) confirmed that TGF‑β1 induced EMT in HTB9 and T24 cells at the protein and mRNA levels. The expression levels of the miR‑200 family were determined by qPCR, which indicated that TGF‑β1 treatment significantly reduced the expression of miR‑200b. Bioinformatic analysis indicated that MMP‑16 may be the target of miR‑200b. Reporter luciferase assay confirmed that MMP‑16 is a direct downstream functional target of miR‑200b. A Matrigel migration assay demonstrated that miR‑200b overexpression inhibited the migration of bladder cancer cells. In summary, the current study demonstrated that exogenous TGF‑β1 leads to the induction of EMT and the downregulation of miR‑200b in bladder cancer cells. To the best of our knowledge, this is the first evidence that MMP‑16 is a direct target of miR‑200b.
Molecular Medicine Reports 07/2014; 10(3). DOI:10.3892/mmr.2014.2366 · 1.48 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Objective
To prospectively study the surgical strategies and clinical efficacy of laparoendoscopic single-site (LESS) inguinal lymphadenectomy compared with conventional endoscopic inguinal lymphadenectomy for the management of inguinal nodes.Patients and MethodsA total of 12 patients with squamous cell carcinoma of the penis who underwent penectomy between February and July 2013 were enrolled in the study. All 12 patients underwent bilateral inguinal lymphadenectomy (LESS inguinal lymphadenectomy in one limb and conventional endoscopic inguinal lymphadenectomy in the other) with preservation of the saphenous vein. All lymphatic tissue in the boundaries of the adductor longus muscle (medially), the sartorius muscle (laterally), 2 cm above the inguinal ligament (superiorly), the Scarpa fascia (superficially) and femoral vessels (deeply) was removed in both surgical techniques. All 24 procedures were performed by one experienced surgeon.ResultsAll 24 procedures (12 LESS and 12 conventional endoscopic inguinal lymphadenectomies) were completed successfully without conversion to open surgery. For LESS inguinal lymphadenectomy and conventional endoscopic inguinal lymphadenectomy groups, the mean ± sd operating time was 94.6 ± 14.8 min and 90.8 ± 10.6 min, respectively (P = 0.145). No significant differences in the incidence of postoperative complications (skin-related problems, hecatomb, lower extremity oedema, lymphatic complications and overall complications) were noted between the two groups (P > 0.05). No lower extremity oedema occurred in any limbs of the two groups. No significant differences were observed in either lymph node clearance rate or detection rate of histologically positive lymph nodes (P > 0.05). The patient satisfaction rate with scar appearance and cosmetic results was significantly better in the LESS inguinal lymphadenectomy group than in the conventional endoscopic inguinal lymphadenectomy group of (75 vs 25%; P = 0.039).Conclusions
This preliminary study suggests that both LESS inguinal lymphadenectomy and conventional endoscopic inguinal lymphadenectomy are safe and feasible procedures for inguinal lymphadenectomy. Preservation of the saphenous vein during LESS inguinal lymphadenectomy/conventional endoscopic inguinal lymphadenectomy can effectively reduce the incidence of postoperative lower extremity oedema. LESS inguinal lymphadenectomy seems to provide better cosmetic results than conventional endoscopic inguinal lymphadenectomy.
BJU International 06/2014; 115(4). DOI:10.1111/bju.12838 · 3.13 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: This study was designed to establish an individualized selection strategy for the two most common types of ureteroenteric anastomotic techniques (Bricker and Wallace anastomosis) used in ileal conduit (IC) diversion.
Patients who underwent IC diversion after radical cystectomy for transitional cell carcinoma between January 2009 and December 2011 were prospectively collected. The choice of anastomosis type (Bricker vs. Wallace) was successively based on tumor characteristics, ureteral anomalies, and ureteral length after retrosigmoidal tunneling.
Ninety-nine patients were enrolled in the final study. Fifty-three patients underwent Bricker anastomosis, and 46 underwent Wallace anastomosis. Ureteral stricture developed in 6 (6.1 %) patients and the overall stricture rate for all ureters was 3.1 % (6/196). Strictures occurred at an average of 13.3 months after surgery and were predominately located in the left ureter (66.7 %, 4/6). The difference in the ureter stricture rates between the two groups was not statistically significant: 3.8 % (4/104) and 2.2 % (2/92) for Bricker and Wallace, respectively (p = 0.686). There were no significant differences in age, sex, body mass index (BMI), prevalence of pelvic radiation therapy, length of stay, follow-up time, or time to stricture between the two techniques. Patients in whom stricture developed had a significantly higher mean BMI compared with those without stricture (25.2 vs. 23.3 kg/m(2), respectively; p = 0.008).
Our preliminary outcomes demonstrate that this selection strategy of Bricker vs. Wallace anastomosis seems to be clinically reliable, providing an acceptable low ureteral stricture rate of 3.1 %. However, the potential advantage for oncologic control of this strategy is needed to further confirm.
Annals of Surgical Oncology 03/2014; DOI:10.1245/s10434-014-3591-z · 3.94 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Objective: To evaluate the safety, efficacy, and potential advantages of off-clamp partial nephrectomy (OFF-PN) compared with on-clamp partial nephrectomy (ON-PN). Methods: Relevant studies comparing the safety and efficacy of OFF-PN to ON-PN were identified through a literature search using MEDLINE, EMBASE, and the Cochrane Library. The outcome measures included baseline characteristics, primary outcomes, and secondary outcomes. Results: Ten retrospective studies (728 cases and 1267 controls) were included. No significant differences between the two groups were detected for any of the baseline variables (age: p = 0.19; sex: p = 0.49; BMI: p = 0.29; tumour size: p = 0.44, pre-eGFR: p = 0.78) except for tumour location (p < 0.001). The OFF-PN group had a higher blood transfusion rate (odds ratio [OR] 1.54, 95% confidence interval [CI] 10.7-2.21, p = 0.02), a lower postoperative complication rate (OR 0.61, 95% CI 0.44-0.83, p = 0.002), and a lower positive margin rate (OR 0.49, 95% CI 0.26-0.90, p = 0.02) than ON-PN. OFF-PN offered a better preservation of renal function than ON-PN (p = 0.005). No significant differences were detected between the two groups in other outcomes of interest. In sensitivity analysis, there was no change in the significance of any of the outcomes except for postoperative complication rate (OR 0.91, 95% CI 0.53-1.5, p = 0.73) and positive margin rate (OR 0.55, 95% CI 0.25-1.23, p = 0.15). Conclusions: This meta-analysis suggests that with appropriate patient selection, OFF-PN offer comparable perioperative safety, equivalent oncologic outcomes, and superior long-term renal function preservation when compared to ON-PN for renal cell carcinoma. Given the inherent limitations of the included studies, future well-designed randomized controlled trials (RCT) are required to confirm our findings.
Journal of endourology / Endourological Society 11/2013; 28(5). DOI:10.1089/end.2013.0562 · 2.10 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: To evaluate the role of two different approaches to perform laparoscopic RPLND: transperitoneal laparoscopic retroperitoneal lymph node dissection (TL-RPLND) and extraperitoneal laparoscopic retroperitoneal lymph node dissection (EL-RPLND).
Between February 2003 and April 2013, 39 patients with nonseminomatous germ cell testicular tumors were treated by RPLND in our center. Twenty-one patients had TL-RPLND, and 18 patients had EL-RPLND. We performed a comprehensive retrospective study comparing TL-RPLND and EL-RPLND. Certain parameters, including operative time, estimated blood loss, perioperative complications, resected lymph nodes, postoperative intestinal function recovery time, ejaculation, and postoperative tumor markers, were abstracted and compared.
In the EL-RPLND and TL-RPLND groups, the operation times were 178 ± 31 and 207 ± 25 min; the amounts of estimated blood loss were 87 ± 26 and 111 ± 21 ml; the postoperative intestinal function recovery times were 1.2 ± 0.7 and 2.4 ± 0.6 days; the postoperative hospital stays were 5.8 ± 1.1 and 5.5 ± 1.4 days; and the numbers of resected lymph nodes were 16.2 ± 1.5 and 15.8 ± 1.6, respectively. No conversion from laparoscopic to open surgery occurred. No patient in either group received an intraoperative blood transfusion. Overall, two patients developed postoperative fever, and one developed abdominal distension. After a median follow-up of 45 months, no regional relapse or metastases occurred, but 4 patients at clinical stage II were treated successfully by three cycles of platinum-based postoperative chemotherapy. Currently, all patients show no evidence of disease.
Our results demonstrate that EL-RPLND was superior to the transperitoneal approach in terms of the operation time, estimated blood loss, and postoperative intestinal function recovery time, whereas no differences were observed in the number of lymph nodes resected. EL-RPLND was demonstrated to be safe and feasible, with satisfactory clinical outcomes when performed by experienced laparoscopic surgeons. Larger cohorts of patients with longer term follow-up are needed for further studies to determine the role of different approaches to L-RPLND.
International Urology and Nephrology 09/2013; 46(2). DOI:10.1007/s11255-013-0547-3 · 1.29 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: OBJECTIVE: To assess long-term effects of biofeedback training on pubertal chronic prostatitis (CP). METHODS: Pubertal CP patients received 12-week intensive biofeedback training and were divided into two groups: group 1 received further monthly training ≥24 (26-36) months; group 2 received further monthly training <24 (13-23) months. National Institutes of Health-CP Symptom Index (NIH-CPSI) scores, maximum urinary flow rate (Qmax) and postvoid residual urine volume (PVR) were recorded monthly. RESULTS: Total NIH-CPSI scores decreased significantly in group 1 (n = 10; mean age ± SD 16.5 ± 1.1 years) together with all subdomain scores (pain, urination, life impact). Total NIH-CPSI scores increased significantly in group 2 (n = 12; mean age ± SD 16.3 ± 1.2 years) at 30 and 36 months, and were significantly different from group 1 at these time points. Urination and life-impact scores increased significantly and Qmax decreased significantly in group 2 at 30 and 36 months. PVR was unchanged in either group. CONCLUSIONS: Twelve-week intensive biofeedback training requires lengthy consolidation sessions to achieve long-term success. Further investigation should assess longer intervals between consolidation sessions, for improving patient compliance and outcome.
The Journal of international medical research 02/2013; 41(2). DOI:10.1177/0300060513477582 · 1.10 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Objective: To study whether TGF-α possesses similar EGF effect of enforcing neuroendocrine differentiation (NED) in prostate cancer cell line DU145 and determine the influence of NED induced by TGF-α on chemoresistance. Methods: DU145 cells were divided into 3 groups: a group with 2% FBS, a group with 2%FBS+TGF-α 5 ng/mL and a group with 2%FBS+TGF-α 10 ng/mL. Morphological change in DU145 cells was observed after TGF-α treatment. Expression levels of NSE mRNA were detected with real time RT-PCR. Western blot was used to detect the expression levels of protein NSE, P-gp, MRP1 and Bcl-2. Cell cycles of DU145 cells in the 3 groups were examined with flow cytometry. MTT assay was used to evaluate the influence of TGF-α in chemoresistance. Results: Compared with DU145 cells cultured with 2% FBS, cells treated with 2% FBS+TGF-α were pleomorphic and pseudopodia extended. The expression level of NSE mRNA upregulated to (3.6±0.5) folds (P<0.05) and (10.1±0.1) folds (P<0.01). Western blot showed that the expression levels of protein NSE, Bcl-2, and MRP1 increased after treatment with different concentrations of TGF-α; P-gp was not detected. The proportion of DU145 cells in phase G1 decreased; proportions of cells in phase S and phase G2/M were increased after TGF-α treatment (5 μg/mL). At the same time, chemoresistance of DU145 cells to cisplatin increased. Conclusion: TGF-α can increase NED in DU145 cells and enforce the chemoresistance to cisplatin.
Zhong nan da xue xue bao. Yi xue ban = Journal of Central South University. Medical sciences 02/2013; 38(2):142-7. DOI:10.3969/j.issn.1672-7347.2013.02.006
[Show abstract][Hide abstract] ABSTRACT: It is challenging to treat renal stones in patients with scoliosis. The present study was designed to study the safety and efficacy of minimally invasive percutaneous nephrolithotomy (mPCNL, 18 Fr) and standard tract PCNL (24 Fr) in patients with scoliosis. Twenty cases treated with mPCNL and 18 cases with standard tract PCNL were included in the present study. Laboratory data included preoperative routine complete blood count, serum creatinine, urine bacterial culture, etc. KUB, intravenous urography or CT scanning was done. Fifteen had lumbar and 23 had thoracolumbar scoliosis. Pulmonary function test was performed in all cases. Demographic and clinical details, operative characteristics and complications were studied and compared between two groups retrospectively. The stone burdens of two groups were averagely 754.4 and 816.2 mm(2), respectively (P = 0.194). Pulmonary function test indicated that 18 (47 %) out of 38 patients had decreased function for surgery and anesthesia. The stone clearance rates were 55 and 67 %, respectively, after the first session (P = 0.522). The requirements of auxiliary treatments including second-look PCNL procedure or SWL (shock wave lithotripsy) were not significantly different for both groups. All patients from both groups achieved final stone clearance after auxiliary treatments. Complications of urinary collecting system injury or fever were observed in one and two cases in each group, respectively, (P = 0.548). There were no injuries to neighboring organs or pneumothorax. The requirement of blood transfusion for four cases in mPCNL group and three cases in the standard tract PCNL group, respectively, indicated no significant difference between two groups (P = 0.999). We are able to successfully and safely perform both mPCNL and standard tract PCNL in patients with scoliosis in our hospital. Compared with mPCNL, standard tract PCNL is even more efficient due to its shorter operative time.
[Show abstract][Hide abstract] ABSTRACT: To explore the anti-tumor effects of asiatic moonseed rhizome extraction-dauricine on bladder cancer EJ cell strain, prostate cancer PC-3Mcell strain and primary cell culture system.
The main effective component-phenolic alkaloids ofMenispermum dauricum was extracted and separated from asiatic moonseed rhizome by chemical method. MTT method was used to detect dauricine anti-tumor effect.
Dauricine had an obvious proliferation inhibition effect on the main tumor cells in urinary system. The minimum drug sensitivity concentration was between 3.81-5.15 μg/mL, and the inhibition ratio increased with the increase of concentration.
Dauricine, the main effective component extracted from asiatic moonseed rhizome, had a good inhibition effect on tumor cells in urinary system. At the same time, Dauricine has certain inhibition effects on the primary cultured tumor cell.
Asian Pacific Journal of Tropical Medicine 12/2012; 5(12):973-6. DOI:10.1016/S1995-7645(12)60185-0 · 0.93 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Prostate cancer metastasis is often associated with poor prognosis. The molecular coupling of the adaptor protein Crk to the docking protein p130(Cas) serves as a switch that regulates cell migration in several invasive cancer cells and Ack appears to act upstream of CrkII to modulate the cell motility. However, the precise role of Ack, Crk and p130(Cas) complex in prostate cancer migration remains unknown. In this study we examined the expression of Crk and p130(Cas) in prostate cancer cell lines, and found that CrkI and p130(Cas) protein level was higher in highly invasive PC-3M and PC-3 cell lines than in moderately invasive DU-145 cells. Upon shRNA mediated knockdown of CrkI and p130(Cas) in PC-3M cells, cell migration and invasion were significantly inhibited as analyzed by wound healing assay and transwell invasion assay. Furthermore, co-immunoprecipitation assay showed that p130(Cas) interacted with CrkI in PC-3M cells and the stability of p130(Cas) and CrkI depended on each other. AckI interacted with both CrkI and p130(Cas) and the interaction of AckI with CrkI seemed to be independent of p130(Cas) . Taken together, our results demonstrate the high expression of CrkI and p130(Cas) in invasive prostate cancer cells and the important role of CrkI/p130(Cas) complex in the migration and invasion of prostate cancer cells. These data suggest that CrkI/p130(Cas) could be exploited as potential molecular therapeutic target for prostate cancer metastasis.
Cell Biochemistry and Function 12/2011; 29(8):625-9. DOI:10.1002/cbf.1797 · 2.13 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Vascular endothelial growth factor-C (VEGF-C) has been found to be significantly associated with lymphangiogenesis and regional lymph node metastasis in various human tumors. The present work was aimed to explore the role of VEGF-C in malignant progression of human bladder cancer T24 cell line. First, the expression of VEGF-C in T24 cells was detected by western blotting. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was employed to measure the cellular proliferation after treatment with various concentrations of recombinant human VEGF-C (rhVEGF-C). Then, lentivirus vector-based RNA interference (RNAi) was used to inhibit VEGF-C expression of T24 cells. The alterations of T24 cells regarding proliferation, invasiveness, and the apoptosis induced by mitomycin C (MMC) were evaluated. The results showed that the proliferation rate of T24 cells rose from 27.3% to 65.0%, with increasing rhVEGF-C concentration. T24 cells stably transfected with VEGF-C small interference RNA showed 85% reduction in VEGF-C mRNA expression (p < 0.05). The VEGF-C protein level was significantly downregulated (p < 0.05) and the growth and invasiveness were also inhibited (p < 0.05) compared with the control group. Further, the inhibition of VEGF-C expression markedly enhanced the apoptosis of T24 cells induced by MMC (p < 0.05). These were associated with the decreased ratio of Bcl-2/Bax, activation of Caspase-3, decreased expression of MMP-9, as well as the downregulation of phosphorylated p38 MAPK and Akt. The present study suggests that VEGF-C can enhance the proliferation and invasiveness of bladder cancer T24 cells, which is due to suppression of apoptosis and facilitation of migration, accompanied with upregulation of p38 MAPK and Akt phosphorylation. RNAi targeting VEGF-C could effectively suppress malignant progression and enhance chemosensitivity of T24 cells. Thus, inhibition of VEGF-C expression is a potential and promising therapeutic strategy for bladder cancer.
[Show abstract][Hide abstract] ABSTRACT: Icaritin has selective estrogen receptor (ER) modulating activity. ERs are expressed in the prostate stroma, and estrogens have an important role in the pathology of benign prostatic hyperplasia (BPH). However, the impact of icaritin on BPH was not studied. Human prostatic smooth muscle cells (PSMCs) were treated with 0-100 microM icaritin, also using 10 microM ICI182780 as a specific ER antagonist. The effects on cell growth and apoptosis were determined by cell counting and sandwich-enzyme-immunoassay. Western blotting was employed to illustrate the possible mechanisms. Cell growth was strongly inhibited by icaritin, and this was accompanied by an augmented apoptosis. Few changes in icaritin-induced growth inhibition and apoptosis were observed after pretreatment in the presence of ICI182780. Consistent with growth inhibition and apoptosis induction, icaritin decreased cyclin D1 and CDK4 expression and increased Bax/Bcl-2 ratio in human PSMCs. Furthermore, icaritin induced sustained phosphorylation of extracellular signal-regulated kinase (ERK) in human PSMCs. PD98059, a specific ERK inhibitor, blocked the activation of ERK by icaritin and abolished the icaritin-induced growth inhibition and apoptosis. The results indicate that icaritin reduces growth and induces apoptosis in human PSMCs via ERK signaling pathway without involvement of ERs.