Oncogenic potential of macrophage­capping protein in clear cell renal cell carcinoma.

Macrophage­capping protein (CapG) is a newly characterized oncogene involved in several types of cancer. However, the expression patterns and biological mechanisms of CapG in clear cell renal cell carcinoma (ccRCC) are unclear. The present study aimed to investigate the roles of CapG in the prognosis, proliferation and metastasis of ccRCC. In the present study, the expression of CapG was analyzed by western blotting in 24 paired ccRCC and adjacent normal tissue samples. Another 152 tissue samples from 152 patients with ccRCC were examined by immunohistochemistry. Compared with normal tissue, CapG expression was significantly increased in ccRCC tissue, and high CapG expression was associated with advanced tumor stage, histological grade, lymph node metastasis, and poor overall survival. Moreover, CapG was an independent predictor of survival. Lentivirus­mediated CapG knockdown significantly inhibited 786­O cell proliferation, migration, and invasion, induced cell cycle arrest at the G2/M phase, and increased apoptosis in vitro. Microarray analysis indicated that RAC, CDC42 and ERK/MAPK signaling were disrupted by CapG knockdown in 786­O cells. In conclusion, the present findings indicate that CapG plays an oncogenic role in ccRCC and may represent a potential therapeutic target for this disease.

[Infiltrating mast cells promote neuroendocrine differentiation and increase docetaxel resistance of prostate cancer cells by up-regulating p21].

OBJECTIVE: To investigate the effect of infiltrating mast cells on neuroendocrine differentiation (NED) and docetaxel sensitivity of prostate cancer (PCa) cells in vitro. METHODS: Human PCa cell lines (LNCaP and C4-2) were co-cultured with human mast cell line (HMC-1) in Transwell chambers. Androgen receptor (AR) was silenced in C4-2 cells using sh-AR lentivirus, and p21 was knocked down and overexpressed by transfecting C4-2 cells with pLKO.1-sh-p21 and pCMV-p21, respectively. The morphological changes of LNCaP and C4-2 cells were observed. MTT assay and colony formation assay were used to assess the proliferation of LNCaP and C4-2 cells. CCK8 assay was used to detect the cell viability of C4-2 cells following docetaxel trreatment. RT-qPCR and Western blotting were performed to determine the mRNA and protein expressions of neuroendocrine markers, AR and p21 in the cells. RESULTS: Co-culture with HMC-1 cells enhanced the neuroendocrine phenotypes, inhibited the proliferation and up-regulated the expression of p21 in LNCaP and C4-2 cells. P21 positively regulated NED through a non-AR-dependent signaling pathway, while p21 knockdown partially reversed NED promoted by the mast cells. PCa cells co-cultured with HMC-1 cells showed increased resistance to docetaxel, and silencing p21 partially reversed docetaxel resistance in PCa cells. CONCLUSION: Infiltrating mast cells up-regulates p21 to promote NED and increase docetaxel resistance in PCa cells in vitro.

[Quantitative and comparative proteomics analysis in clear cell renal cell carcinoma and adjacent noncancerous tissues by 2-D DIGE].

OBJECTIVE: To identify specific protein markers for renal cell carcinoma detection and diagnosis, as well as develop new potential therapeutic targets of the disease. METHODS: We used two-dimensional difference in-gel electrophoresis (2-D DIGE) technique conjunction with mass spectrometry (MS) for the identification of significant differentially expressed proteins between 15cases of paired clear cell renal cell carcinoma (ccRCC) and adjacent normal renal tissues. The protein spots were considered as differentially expressed if a 1.5-fold altered expression level was observed (Student's t test, P value<0.05). RESULTS: Of the 27 differentially expressed protein spots, 26 proteins were successfully identified. 11 proteins up-regulated in renal cell carcinoma,15 proteins down-regulated. Among them Short/branched chain specific acyl-CoA dehydrogenase, mitochondrial (ACDSB), Aldose 1-epimerase (GALM), Peroxiredoxin-4 (PRDX4), Macrophage-capping protein (CAPG), Beta-defensin 107 (D107A), Microfibril-associated glycoprotein 4 (MFAP4) were first time screening as new differential expressed proteins by protomic study in renal cell carcinoma. CONCLUSIONS: 2-D DIGE is a useful technique for screening and analysis differential expressed proteins in renal cell carcinoma. These new differently expressed proteins may be useful for development new molecular markers for the tumor.

Saikosaponin-d: A potential chemotherapeutics in castration resistant prostate cancer by suppressing cancer metastases and cancer stem cell phenotypes.

Androgen deprivation therapy is the gold standard regimen for advanced Prostate cancer (PCa) patients, nevertheless, patients eventually develop into castration-resistant prostate cancer (CRPC). Currently only a few chemotherapeutics are available for CRPC. Therefore, it is critical for identifying a new drug. In this study, we will explore a new agent, Saikosaponin-d (SSd), for CRPC therapy based on its mechanism of action. DU145 and CWR22Rv1 cells representing CRPC were employed in this study. A series of cell, biochemical, and molecular biologic assays such as Immunofluorescence, Zymography, Sphere formation, Colony formation, and MTT were used. Finally, we find SSd can significantly inhibit the growth of PCa cells in both dose- and time-dependent and suppress the colony formation during a long-term drug administration, it also can inhibit their migration and invasion abilities, which was accompanied by reverse the epithelial-mesenchymal transition (EMT) and suppress MMP2/9 expression as well as activities. Furthermore, SSd can suppress cancer stem cell (CSC) phenotypes such as self-renewal ability. Mechanistically, SSd blocks Wnt/ß-catenin signaling pathway by decreasing GSK3ß phosphorylation to affect EMT and CSC. These findings demonstrate the mechanism of anti-cancer activity of SSd in targeting EMT and CSC, suggesting SSd can be a potent agent for CRPC therapy.

[Value of MN/CAIX in the diagnosis of renal cell carcinoma].

OBJECTIVE: To investigate the expression of MN/CAIX in patients with renal cell carcinoma (RCC) and assess the value of MN/CAIX in the diagnosis of RCC. METHODS: RT-PCR was employed to detect MN/CAIX mRNA in the carcinoma tissue and peripheral blood of 62 patients with RCC, using normal renal tissue and peripheral blood sample from 32 patients without RCC as control. Immunohistochemistry was used to detect MN/CAIX protein in the tissue specimens of clear cell RCC (n=36), non-clear cell renal neoplasm (n=17) and normal kidney (n=16). RESULTS: The positivity rate of MN/CAIX mRNA was 82.3% (51/62) in renal carcinoma tissues and 54.8% (34/62) in the peripheral blood from patients with RCC, significantly higher than the rates in the control cases (P<0.05). In cases of clear cell renal cell carcinoma, the positivity rate of MN/CAIX mRNA was 98% (49/50) in the carcinoma tissues and 66% (33/50) in the peripheral blood, significantly higher than the rates in cases of non-clear cell type of RCC (P<0.05). Immunohistochemistry showed a significantly higher positivity rate of MN/CAIX protein in clear cell RCC tissues [97.2% (35/36)] than in non-clear cell renal neoplasm and normal renal tissues (P<0.05). CONCLUSION: MN/CAIX is specifically overexpressed in RCC, especially in clear cell RCC, suggesting its potential in the diagnosis and prognostic and therapeutic evaluation of RCC.

[Synchronous squamous cell carcinoma of the renal pelvis and squamous cell carcinoma of the ureter: report of two cases and review of literature].

OBJECTIVE: To study the clinicopathological characteristics of synchronous squamous cell carcinoma (SCC) of the renal pelvis and SCC of the ureter. METHODS: The clinical data of two cases of synchronous SCC of the renal pelvis and SCC of the ureter were retrospectively reviewed and analyzed. In case 1, a 68-year-old man with hematuria for a month, imaging modalities revealed a right renal pelvis tumor and a right distal ureter tumor. The patient underwent nephroureterectomy and excision of the bladder cuff. Case 2, a 60-year-old man with the complaint of lower abdominal pain and left flank pain for a month, was diagnosed as left distal ureteral stone in another hospital. Ureterolithotomy was performed and a ureteral tumor was found at the lower site of the stone intraoperatively. The pathological report demonstrated SCC, and the patient was transferred to our hospital for further treatment. We found a left renal mass invading the left hemicolon during surgery, and nephroureterectomy was performed with a bladder cuff excision, left hemicolon resection, and also complete lymph node dissection. Neither of patients received adjuvant radiotherapy/chemotherapy. RESULTS: Moderately differentiated SCC was reported in both of renal pelvis and ureter in case 1 and the tumor invaded the subepithelial connective tissue in the renal pelvis and superficial muscle in the ureter. In case 2, moderately differentiated SCC of the left renal pelvis with colon metastasis and poorly differentiated SCC of the ureter was reported with two retroperitoneal lymph node metastases. The two patients died from tumor recurrence and metastasis 5 and 6 months after the surgery, respectively. CONCLUSION: Synchronous SCC of the renal pelvis and SCC of the ureter are rare and has high likeliness of early recurrence and metastasis, often with poor prognosis.

[Detection of Skp2 and p27kip1 expression in human renal cell carcinoma using tissue chip technique].

OBJECTIVE: To detect the expression of skp2 and p27kip1 in human renal cell carcinoma (RCC) using tissue chip technique, and evaluate the relationship between the proteins and the biological behavior of RCC. METHODS: Tissue chip technique and immunohistochemical SP method was used to detect the expression of skp2 and p27kip1 in normal and tumor tissues. RESULTS: The positivity rate of Skp2 in RCC was significantly higher than that in normal renal tissues (P=0.025). The positivity rate of Skp2 expression in RCC was significantly correlated to poor differentiation of the tumor (P=0.002), and was not associated with the patients gender, age, tumor size, lymph node metastasis and stages of RCC (P>0.05). The positivity rate of p27kip1 in RCC was significantly lower than that in normal renal tissues (P=0.007). The positivity rate of p27kip1 expression was inversely correlated to the malignancy and stage of RCC (P<0.05), but not with the patients' age, gender, lymph node metastasis and tumor size (P>0.05). An inverse correlation was noted between Skp2 and p27kip1 expressions (r= -0.273, P=0.014). CONCLUSION: Overexpression of Skp2 protein may lead to decreased p27kip1 level in RCC, indicating its involvement in the carcinogenesis and development of RCC.

[Expression of human telomerase reverse transcriptase in renal cell carcinoma and its clinical significance].

OBJECTIVE: To investigate the expression of human telomerase reverse transcriptase (hTERT) in renal cell carcinoma (RCC) and its clinical significance. METHODS: The expression levels of hTERT mRNA and protein were detected using RT-RCR and Western blotting in 45 RCC tissues, 45 adjacent tissues and 786-0 cell line, and the associations of hTERT expression with the tumor size, clinical stage, pathological type and grade were evaluated. RESULTS: hTERT mRNA and protein was expressed at significantly higher levels in RCC tissues than in the adjacent tissues (P=0.000), and no correlation of hTERT expression was found with the tumor size, clinical stage, pathological type or grade. CONCLUSION: hTERT might serve as a diagnostic and prognostic marker for RCC, and also shed light on the new clues for gene therapy of RCC.

[Fusion expression of human renal cell carcinoma-associated antigen G250/MN/CA IX in prokaryotic expression system].

OBJECTIVE: To achieve high expression of human renal cell carcinoma-associated antigen G250 in Escherichia coli. METHODS: The gene fragments encoding the protein obtained by PCR was cloned into prokaryotic expression vector pET32a(+) and expressed in E. coli Rosseta. The immunogenicity of the recombinant protein was evaluated by Western blotting. RESULTS: The plasmid pET32a(+)/G250 was constructed and expressed in E. coli Rosseta successfully. Western blot analysis showed that the recombinant protein could be specifically recognized by monoclonal antibody M75. CONCLUSION: Efficient G250 expression is achieved in prokaryotic expression system, which may facilitate further functional study of the protein and its monoclonal antibody preparation.

[Expressions of hypoxia inducible factor-1alpha and vascular endothelial growth factor in human renal cell carcinoma].

OBJECTIVE: To explore the expression of hypoxia inducible factor-1alpha (HIF-1alpha) and vascular endothelial growth factor (VEGF) in renal cell carcinoma (RCC) and their relationship. METHODS: The expression of HIF-1alphaand VEGF was examined in 42 patients with renal cell carcinoma by SABC methods of immunohistochemistry. RESULTS: The positivity rate of HIF-1alpha was 57.1% in the RCC tissues, and there was no expression in the tissue adjacent to the tumor and normal tissues. The positivity rate of VEGF was 61.9% in the tumor tissues, with significant difference from the control tissues (P<0.05). The expression levels of HIF-1alpha and VEGF were significantly higher in metastatic RCC than in non-metastatic RCC (P<0.01) tissues, and there was a significant correlation between HIF-1alpha and VEGF expressions (Kappa=0.41, P<0.01). CONCLUSIONS: HIF-1alpha is highly expressed in renal cell carcinoma tissues in close correlation with VEGF. HIF-1alpha and VEGF may serve as important indicators to evaluate the biological behaviors of RCC such as metastasis and prognosis. HIF-1alpha is an important regulator of VEGF.

[Nephron-sparing surgery for small renal cell carcinoma: clinical analysis of 21 cases].

OBJECTIVE: To evaluate the clinical effects of nephron-sparing surgery in patients with early-stage small renal cell carcinoma. METHODS: Nephron-sparing surgery was performed in 21 patients with renal cell carcinoma including 1 with solitary kidney, 3 with unilateral tumor and contralateral renal compromise, and 17 with unilateral tumor and normal contralateral kidney. The diameter of the tumors ranged from 1.5 to 6.0 cm, with a mean of 2.8 cm. The tumor diameter in 17 patients with normal contralateral kidney was less than 4 cm (mean 2.5 cm) and the average diameter in 4 patients with contralateral renal compromise was 4.2 cm. Sixteen cases were in stage T(1), 4 in stage T(2), and 1 in stage T(3). Of the 21 patients, 4 underwent tumor enucleation, 10 polar nephrectomy and 7 wedge resection. RESULTS: All patients were followed up for an average of 40.8 months (7 to 66 months). One patient suffered a right lung and mediastinum metastasis 3 years after the surgery later and 1 with chronic glomerulonephritis required dialysis 27 months after the operation. No surgical complication or local recurrence were found in other patients. CONCLUSION: As a safe and effective therapy for early-stage small renal cell carcinoma, nephron-sparing surgery can be considered as the gold-standard therapy for patients with lesions less than 4 cm in T(1) and T(2) stages of localized unilateral tumor with normal contralateral kidney.

[MN/CAIX gene expression in renal cell carcinoma detected by RT-PCR].

OBJECTIVE: To evaluate the reliability of reverse transcriptional (RT)-PCR in detecting MN/CAIX gene expression in renal cell carcinoma (RCC). METHODS: MN/CAIX gene expression was detected in 5 RCC tissues and 5 normal renal tissues. The primers of MN/CAIX gene were designed for a two-step RT-PCR, and the products examined by 2% agarose gel electrophoresis and identified by sequence analysis. RESULT: Corresponding bands of MN/CAIX gene could be seen in RT-PCR products of cases RCC tissue from 4 but not in normal renal tissues. CONCLUSION: RT-PCR is reliable for detecting the expression of MN/CAIX gene in RCC.

Preparation and characterization of polybutylcyanoacrylate magnetic nanoparticles.

OBJECTIVE: To Prepare and characterize polybutylcyanoacrylate (PBCA) magnetic nanoparticles. METHODS: The suspension of PBCA magnetic nanoparticles was prepared by emulsion polymerization method. The particle size, size distribution, polydispersity and other parameters were determined by photon correlation spectroscopy and transmission electron microscopy, and the iron content in the nanoparticles by ultraviolet spectroscopy. RESULTS: The data of the particle size and size distribution of aqueous decanoic acid stabilized Fe3O4 magnetic fluids were acquired, and the influences of concentrations of butylcyanoacrylate (BCA) and Dextran-70 on the particle size and size distribution of PBCA magnetic nanoparticles were characterized. CONCLUSION: We have for the first time successfully prepared stable suspension of PBCA magnetic nanoparticles by emulsion polymerization method at pH 6.3-6.4 in aqueous medium.

Treatment of the posterior urethral stricture: experience with 13 cases.

OBJECTIVE: To review our experience with the treatment of posterior urethral stricture. METHOD: A elaborated retrospective analysis of the treatment of 13 cases of posterior urethral stricture was conducted with the typical cases. RESULT: Good therapeutic results were achieved in these cases after treatment with cryoablation, electrotome or open surgery. CONCLUSION: Proper initial treatment, adequate surgical approach, anti-infection measures and postoperative urethral dilatation are the key elements in successful treatment of posterior urethral stricture.

[Detection of carbonic anhydrase IX gene expression in renal cell carcinoma with reverse transcription-PCR].

OBJECTIVE: To evaluate the reliability of reverse transcription (RT)-PCR in detecting the expression of the carbonic anhydrase IX (MN/CAIX) gene in renal cell carcinoma (RCC). METHODS: MN/CAIX was examined in 5 cases of renal cell carcinoma tissues and 5 normal renal tissues. Primers specific for MN/CAIX was designed and two-step RT-PCR of the extracted total RNA performed. The products was examined by 2% agarose electrophoresis and identified by sequence analysis. RESULTS: Specific bands of MN/CAIX were found in RT-PCR products of 4 case of RCC tissues, whereas could not be seen in normal renal tissues. CONCLUSION: RT-PCR is reliable for detecting the expression of MN/CAIX gene in RCC.

[Selection of the imaging modalities for diagnosis of renal trauma: experience with 74 cases].

OBJECTIVE: To explore the diagnostic approaches and values of the imaging modalities for traumatic renal injuries. METHODS: The clinical records of 74 cases of renal trauma treated in Nanfang Hospital were retrospectively reviewed to assess the diagnostic value of intravenous urography (IVU), type B ultrasonography and computed tomography (CT). RESULTS: The positivity rates by IVU, type B ultrasonography, and CT were 89% (43/49), 80% (55/68) and 100% (51/51) respectively for the diagnosis of renal trauma. CONCLUSION: IVU is rapid and convenient, ultrasonography less costly and invasive, and CT accurate for diagnosis of renal traumas.