[Ten significantly differentially expressed genes in prostate cancer: Screening and verification].

OBJECTIVE: To screen and verify differentially expressed genes in prostate cancer. METHODS: Using DNA microarray, we screened differentially expressed genes in prostate cancer tissue and its adjacent tissue followed by verification by PCR. RESULTS: A total of 1 444 genes were found to be differentially expressed (differentiation ≥ 1.5-fold; P≤ 0.05) in the prostate cancer tissue, of which 769 (53%) were up-regulated and 675 (47%) down-regulated. Fifty percent of the differentially expressed genes showed a 1.5- to 2-fold differentiation, including 396 up-regulated and 182 down-regulated ones. Additionally, 308 up-regulated and 334 down-regulated genes exhibited a >2- to 5-fold, 46 up-regulated and 78 down-regulated genes a > 5- to 10-fold, and 19 up-regulated and 81 down-regulated genes a > 10-fold differentiation. Verification by subjecting 15 most significantly up-regulated and another 15 most markedly down-regulated genes to quantitative real-time PCR (qRT-PCR) showed that most of the genes had a transcriptional profile similar to that in the microarray data, with a Pearson correction coefficient of 0.83 between the microarray data and qRT-PCR results. Totally, 10 significantly differentially expressed genes were identified. CONCLUSION: DNA microarray analysis provides reliable information on differentially expressed genes in prostate cancer and benign tissues. The 10 significantly differentially expressed genes verified by qRT-PCR could possibly become new bio-markers and specific molecules for tumor identification.

Expression of CD147 is associated with prostate cancer progression.

Novel molecular markers that are associated with prostate cancer (PCa) progression will provide valuable information in the diagnosis and treatment of the disease. Extracellular matrix metalloproteinase inducer (CD147) has been demonstrated to be involved in tumor invasion, metastasis, growth and survival. In our study, we examined whether the expression of CD147 can be used as a prognostic marker for predicting PCa progression. Tissue samples from 240 patients who received radical prostatectomy for PCa were obtained. CD147 expression in these samples was evaluated using immunohistochemical staining with a monoclonal antibody specifically against CD147. Increased expression of CD147 was correlated with higher Gleason scores (GS), positive surgical margin, prostate-specific antigen (PSA) failure, metastasis and reduced overall survival. Both univariate Cox regression analysis and multivariate analysis including competing biological variables demonstrated that increased CD147 expression was associated with increased risk for reduced PSA failure-free, metastasis-free and overall survival. Kaplan-Meier survival curves showed that the CD147 overexpression was a significant predictor for the PSA failure-free, metastasis-free and the overall survival in both pT2 and pT3 PCa patients. More significantly, higher expression of CD147 can serve as an independent prognostic predictor for PSA failure-free survival in PCa patients when they are stratified by GS. Our study results demonstrate the involvement of CD147 in PCa progression and suggest its potential role as an independent predictor of biochemical recurrence, development of metastasis and reduced overall survival in PCa.

Expression and clinical significance of CD147 in genitourinary carcinomas.

BACKGROUND: CD147/extracellular matrix metalloproteinase inducer (EMMPRIN) expressed by tumor cells stimulates peri-tumorous fibroblasts to produce matrix metalloproteinases (MMPs), thus contributing to tumor invasion and metastasis. To assess its suitability as a potential therapeutic target, as well as its association with the clinicopathologic features and the prognosis of patients, the expression of CD147/EMMPRIN in neoplastic tissues of the genitourinary system were analyzed. METHODS: CD147/EMMPRIN expression in 52 patients with renal carcinoma, 58 patients with bladder carcinoma, 101 patients with prostate carcinoma, 17 patients of penis carcinoma, and 17 patients of testis carcinoma were examined by immunostaining on paraffin-embedded tumor specimens using monoclonal antibodies. Then, the association of its expression with clinicopathologic characteristics to the patients' prognosis was analyzed. The RNA interference approach was used to silence CD147/EMMPRIN expression in the human prostate carcinoma cell line LNCAP and human bladder carcinoma cell line J82. The in vitro proliferative ability of CD147/EMMPRIN-deficient cells was determined by a 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide MTT assay. RESULTS: CD147/EMMPRIN was expressed in neoplastic tissues, but not in normal tissues. Positive expression was shown in 42 of 52 (80.77%) of the patients with renal carcinoma, 41 of 58 (70.69%) of the patients with bladder carcinoma, 67 of 101 (66.34%) of the patients with prostate carcinoma, 16 of 17 (94.12%) of the patients with penis carcinoma and testis carcinoma. Positive CD147/EMMPRIN staining was significantly associated with TNM stages and histological subtypes of patients with various urinary carcinomas (P < 0.05). In all five groups, for different expression levels of CD147/EMMPRIN, the patients with a highly positive expression of CD147/EMMPRIN had the poorest prognosis. The siRNA-treated cells exhibited significantly decreased growth ability compared with control cells in vitro. CONCLUSION: These results may assist in defining the suitability of CD147/EMMPRIN as a therapeutic target and as a method for predicting a poor outcome in patients with various urinary carcinomas.

Inhibition of proliferation, invasion, and migration of prostate cancer cells by downregulating elongation factor-1alpha expression.

Overexpression of elongation factor-1alpha (EF-1alpha) has been reported to contribute to the development and progression of various cancers. However, its role in prostate cancer (PCa) still remains poorly understood. In the present study, we investigate the influence of EF-1alpha in Du145, a high-grade metastatic PCa cell line, and demonstrate that EF-1alpha plays an essential role in cellular properties associated with tumor progression, namely cell proliferation, invasion, and migration. In this study, EF-1alpha expression in human PCa cell line Du145 was reduced by RNA interference (RNAi) technology, and the proliferation, invasion, and migration of EF-1alpha-reduced Du145 cells were examined. We also detected an EF-1alpha expression pattern in 20 pairs of primary PCa samples and their corresponding normal tissues. Expression of EF-1alpha was detectable in four PCa cell lines (22RV1, LnCap, Du145, and PC3), indicating its possible role in pathogenesis of PCa. RNAi-mediated knockdown of EF-1alpha expression in Du145 cells, which expressed the highest level of EF-1alpha among four PCa cell lines, led to a decrease in proliferation. Similarly, suppression of EF-1alpha inhibited Du145 cell migration and invasion through a basement membrane substitute. Furthermore, we found that the normal prostate tissues showed a relatively low level of EF-1alpha expression, whereas PCa tissues demonstrated significantly higher expression levels of EF-1alpha (P < 0.001). Taken together, these findings support the hypothesis that EF-1alpha affects multiple processes involved in tumor progression, and identify EF-1alpha as a potential therapeutic target.

CD147 and VEGF expression in advanced renal cell carcinoma and their prognostic value.

AIM: To investigate the clinicopathologic characteristics of extracellular matrix (ECM) metalloproteinase inducer (CD147) and vascular endothelial growth factor (VEGF) expression in advanced renal cell carcinoma (RCC), and to evaluate the clinical significance of these two markers in the prognosis of advanced RCC. METHODS: CD147 and VEGF expression in paraffin-embedded specimens gathered from 53 patients with advanced RCC and 12 healthy controls were detected by the method of immunohistochemistry. The Spearman correlation was calculated between the expression levels of CD147 and VEGF in advanced RCC tissues. The association of CD147 and VEGF expression with the clinicopathologic features and prognosis of advanced RCC was subsequently assessed. RESULTS: CD147 and VEGF were positively expressed in 47/53 (88.7%) and 45/53 (84.9%) of patients with advanced RCC, respectively. Positive expression of CD147 (p= 0.02) and VEGF (p< 0.01) was significantly correlated with TNM stage of advanced RCC. A significant correlation was found between the expression of CD147 and VEGF in advanced RCC (r= 0.629, p= 0.04). Additionally, tumor CD147 and tumor VEGF expressions were significantly associated with the prognosis of advanced RCC patients. The survival rate of the patients with CD147-/VEGF- expression was the lowest (p< 0.01), and conjoined expressions of CD147-/VEGF- and CD147+/VEGF+ were independent prognostic indicators of advanced RCC (both p< 0.01). CONCLUSION: The expression of CD147 or VEGF may be an important feature of advanced RCC. A combined detection of CD147/VEGF coexpression may benefit us in the prediction of the prognosis of advanced RCC.

Real-time quantitative RT-PCR assessment of PIM-1 and hK2 mRNA expression in benign prostate hyperplasia and prostate cancer.

To investigate the expressions of PIM-1 and hK2 mRNA in normal prostate, benign prostatic glandular hyperplasia (BPH), and prostate cancer (PCa), and to explore the association of PIM-1 and hK2 expressions with PCa progression. The samples were harvested from 37 patients with BPH, 23 patients with PCa, and three with normal prostate tissues. Total RNA was extracted from their prostate tissues and analyzed for PIM-1 and hK2 mRNA levels using SYBR green I-based quantitative real-time RT-PCR (QRT-PCR) assays and Southern blot analysis. The differences of gene expressions were calculated based on standard curve. Quantitative expressions of PIM-1 and hK2 mRNA in normal prostate, BPH, and PCa were 1.05 +/- 0.04, 2.57 +/- 0.74, 4.45 +/- 0.63, and 1.02 +/- 0.03, 2.264 +/- 0.46, 5.905 +/- 0.78, respectively. PIM-1 and hK2 were expressed higher in PCa than those in BPH and normal prostate tissues, the differences among which had statistic significance (P < 0.05). Our results support the hypothesis that PIM-1 and hK2 play a significant role in the growth of PCa and the detection of PIM-1 and hK2 mRNA expressions by QRT-PCR provided more reliable and helpful information on diagnosis, treatment, and prognosis of PCa.

[Expression and significance of CD147 protein in prostate cancer].

OBJECTIVE: To investigate the expression of CD147 in prostate cancer and discuss its diagnostic value in prostate cancer. METHODS: The method of immunohistochemical SP was employed to detect the expression of CD147 in 101 cases of prostate cancer, 90 cases of benign prostatic hyperplasia, 36 cases of normal prostate and 15 cases of embryonic prostate by so as to evaluate its clinical significance in the histological classification and prognosis of prostate cancer. RESULTS: The CD147 expression was positively expressed in 67/101 (66.3%) of prostate cancer, 21/90 (23.3%) of benign prostatic hypertrophy, 2/36 (5.6%) of normal prostate and 0/15 (0.0) of embryonic prostate respectively. A positive expression of CD147 was dramatically associated with TNM stage (P < 0.001), the depth of prostatic capsule invasion (P = 0.002) and histological grade (P = 0.006). CONCLUSIONS: The detection of CD147 is helpful to raise the early diagnosis rate of prostate cancer. It will become a tumor marker of reflecting the malignant degree and predicting the prognosis of prostate cancer.

CD147, MMP-1, MMP-2 and MMP-9 protein expression as significant prognostic factors in human prostate cancer.

AIM: CD147 and MMPs have been demonstrated to be involved in tumor invasion and angiogenesis. The aim of this study was to analyze the clinicopathological significance of CD147, MMP-1, MMP-2 and MMP-9 expression in human prostate cancer (PCa) and to evaluate their involvement in the progression of PCa. METHODS: CD147, MMP-1, MMP-2 and MMP-9 expression was assessed in paraffin-embedded specimens collected from 62 cases of PCa and 15 cases of benign prostatic hyperplasia (BPH) by immunohistochemistry. Spearman's correlation was applied to determine possible relationships between CD147, MMP-1, MMP-2 and MMP-9 expression and PCa. The association of CD147 and MMP-2 protein expression with the clinicopathological characteristics and the prognosis of PCa was subsequently assessed. RESULTS: CD147was expressed in 51/62 (82.3%) PCa patients and in 2/15 (13.3%) BPH cases. MMP-1, MMP-2 and MMP-9 expression was significantly higher in PCa tissue than in BPH tissue. Using Spearman analysis, a significant positive correlation between CD147 and MMP-1, MMP-2 and MMP-9 expression was found (p <0.05). CD147 and MMP-2 expression was correlated with TMN grade and Gleason score. Patients with concurrent expression of CD147+ and MMP-2+ had the lowest survival (p <0.01). CONCLUSION: The results suggest that concurrent expression of CD147 and MMP may be an important characteristic of PCa which may help in the prediction of PCa progression.

Extracellular matrix metalloproteinase inducer: a novel poor prognostic marker for human seminomas.

OBJECTIVE: Extracellular matrix metalloproteinase inducer (EMMPRIN) is a glycosylated member of the immunoglobulin superfamily whose function in human seminomas is unknown. We have recently determined that EMMPRIN possesses the ability to stimulate fibroblast and endothelial cell matrix metalloproteinase production, and that its expression was frequently up-regulated in several tumours of the urinary system. Thus, EMMPRIN expression might be associated with the progression of human seminomas. The aim of this study was to investigate whether the presence of EMMPRIN in seminoma tissues might help to predict the patients' prognosis. METHODS: Paraffin-embedded tissues from 65 patients with seminomas and 20 normal testes were processed for immunohistochemical staining using a mouse monoclonal antibody generated against human EMMPRIN, as primary antibody, and a biotinylated goat-anti-mouse IgG, as secondary antibody. In addition, the correlation of EMMPRIN expression with clinicopathologic characteristics and patients' prognosis was also analysed. RESULTS: EMMPRIN was detected in cancerous tissues of 53 patients with seminoma, but not normal testes. Thirty- five patients showed weakly to moderately positive and 18 patients intensely positive expression. Moreover, positive EMMPRIN staining correlated significantly with various clinicopathological factors (increased TNM stage and higher histological differentiation type) as well as decreased tumour-specific survival (log-rank, p=0.02). In particular, EMMPRIN expression was an independent prognosticator as shown by Cox regression analysis (p<0.001). CONCLUSION: EMMPRIN expression in a primary tumour predicts an unfavourable prognosis in human seminoma, suggesting its crucial role in the progression of this tumour.

CK20 and Ki-67 as significant prognostic factors in human bladder carcinoma.

Aberrant expression of CK20 and Ki-67 has been documented in many kinds of primary tumors and has proved useful as an ancillary diagnostic aid for those tumors. The aim of this study was to analyze the expression patterns of CK20 and Ki-67 in human bladder carcinomas (BCa) and to evaluate their clinical significance in the progression of BCa. CK20 and Ki-67 expression in BCa and normal bladder tissues were detected by immunohistochemical staining. The Spearman correlation was calculated between the expression of CK20 and Ki-67 in BCa tissues. The correlation of CK20 and Ki-67 expression with the clinicopathological characteristics and the prognosis of BCa were subsequently assessed. CK20 expression was positively expressed in 103/154 (66.9%) of BCa and 2/30 (6.67%) of normal bladder tissues, respectively. The positive expression rate of Ki-67 in BCa tissues was also significantly higher than those in normal bladder tissues (81.8 vs. 10%, p < 0.01). The Spearman analysis indicated that the expression level of CK20 has a significant positive correlation with that of Ki-67 (rs = 0.86, p = 0.02). Pathologic findings demonstrated that the intensity of CK20 and Ki-67 staining in cancerous tissues was associated significantly with tumor grades (p = 0.03, p < 0.01), distant metastasis (both p < 0.01) and TNM grades (p = 0.01, p = 0.03) of BCa. The progression-free survival of the patients with CK20 (+)/Ki-67 (+) expression was poorest (p < 0.01). The results suggest that the expression of CK20 and Ki-67 may be an important feature of BCa, and the detection of their co-expression may benefit the prediction of BCa prognosis.

Extracellular matrix metalloproteinase inducer expression has an impact on survival in human bladder cancer.

AIM: Extracellular matrix metalloproteinase inducer (EMMPRIN) has been shown to promote tumor invasion and metastasis via stimulating matrix metalloproteinase synthesis in neighboring fibroblasts, to enhance angiogenesis via vascular endothelial growth factor, to induce chemoresistant tumor cells via the production of hyaluronan, and to confer resistance of cancer cells to anoikis through inhibition of Bim. The purpose of this study was to investigate the expression of EMMPRIN in human primary bladder cancer and to evaluate its prognostic value. METHODS: EMMPRIN expression patterns were detected by immunohistochemistry. In order to determine its prognostic value, overall survival (OS) and progression-free survival (PFS) were evaluated using the Kaplan-Meier method, and multivariate analysis was performed using the Cox proportional hazard analysis. RESULTS: Of the 101 cases with bladder cancers, 68 (67.3%) cases were positive for EMMPRIN expression. When categorized into negative vs. positive expression, EMMPRIN was associated with the stage (p=0.006), the grade (p=0.002), carcinoma in situ (p=0.01), the recurrence (p=0.009), the progression (p=0.009), and the death (p=0.01) of patients with bladder cancer. Moreover, positive EMMPRIN expression clearly predicted poorer PFS (p=0.008) and OS (p=0.006). In the multivariate analysis, positive EMMPRIN expression was an independent prognostic factor for PFS (p=0.03) and OS (p=0.03). CONCLUSION: EMMPRIN expression was greater in bladder cancers than in the adjacent normal tissues and may be a useful prognostic marker for patients with bladder cancer.

Pathogen incidence and antibiotic resistance patterns of catheter-associated urinary tract infection in children.

The aim of this study was to characterize the pathogens and their antibiotic susceptibilities in children with catheter-associated urinary tract infection (CAUTI) in order to optimize empirical antibiotic therapy and prophylaxis. from 2001 to 2006, 895 children with an indwelling catheter from 3 hospitals in China were included in this study, of whom 335 (37.4%) had CAUTI. Antimicrobial susceptibility testing of 450 bacterial isolates was performed using the ClSi broth and Kirby-bauer agar dilution methods. Escherichia coli was the most frequently isolated pathogen, followed by Staphylococcus aureus, Staphylococcus epidermidis and Enterococcus spp. E. coli had higher susceptibility to ceftazidime (87.4%), cefuroxime (85.1%) and cefatrizine (76.6%) than to sulfamethoxazole (SMZ) (8.0%), amoxicillin (21.7%), ampicillin (17.1%) and cefazolin (37.7%). Isolates of Klebsiella pneumoniae and Proteus species had similar patterns as E. coli. S. aureus had lower susceptibility to SmZ (6.8%), ampicillin (8.2%), and amoxicillin (24.7%); the trend of S. epidermidis was similar. This study demonstrates that the Gram-negative species are the predominating uropathogens of CAUti in children. it is important to know the bacterial spectrum and the susceptibility patterns to various classes of antibiotic agents to improve empiric antibiotic therapy of children with CAUTI in China.

CD147 expression indicates unfavourable prognosis in prostate cancer.

Extracellular matrix metalloproteinase inducer (EMMPRIN, also named as CD147) is a multifunctional membrane glycoprotein over-expressed in many kinds of human solid tumors. It has been demonstrated to be involved in tumor invasion and angiogenesis. The aim of this study was to analyze the clinicopathological characteristics of the expression of CD147 in human prostate cancer (PCa), and to evaluate its clinical significance in the histologic classification and prognosis of PCa. CD147 protein expression in paraffin-embedded specimens gathered from 62 cases of PCa and 30 cases of benign prostatic hyperplasia (BPH) were detected by the method of immunohistochemistry. The association of CD147 protein expression with the clinicopathological characteristics and with the prognosis of PCa was subsequently assessed. CD147 expression were positively expressed in 51/62 (82.3%) of PCa and 4/30 (13.3%) of BPH cases, respectively. The positive expression rate of CD147 in PCa tissues was significantly higher than that in BPH. The positive expression of CD147 was dramatically associated with TNM grade (p < 0.001), the depth of the prostatic wall invasion (p = 0.008), GLEASON Score (p = 0.001) and Histologic grade (p = 0.001). The patients with CD147 expression were associated with a poor prognosis of PCa (p = 0.01) and the survival rate of the patients with a strong positive expression of CD147 was the lowest (p = 0.01). The results suggest that the expression of CD147 may be an important feature of PCa and the detection of its expression may benefit us in the prediction of the prognosis of PCa.

Hybridization biosensor using di(2,2'-bipyridine)osmium (III) as electrochemical indicator for detection of polymerase chain reaction product of hepatitis B virus DNA.

A novel hepatitis B virus (HBV) DNA biosensor was developed by immobilizing covalently single-stranded HBV DNA fragments to a gold electrode surface via carboxylate ester to link the 3(')-hydroxy end of the DNA with the carboxyl of the thioglycolic acid (TGA) monolayer. A short-stranded HBV DNA fragment (181bp) of known sequence was obtained and amplified by PCR. The surface hybridization of the immobilized single-stranded HBV DNA fragment with its complementary DNA fragment was evidenced by electrochemical methods using [Os(bpy)(2)Cl(2)](+) as a novel electroactive indicator. The formation of double-stranded HBV DNA on the gold electrode resulted in a great increase in the peak currents of [Os(bpy)(2)Cl(2)](+) in comparison with those obtained at a bare or single-stranded HBV DNA-modified electrode. The mismatching experiment indicated that the surface hybridization was specific. The difference between the responses of [Os(bpy)(2)Cl(2)](+) at single-stranded and double-stranded DNA/TGA gold electrodes suggested that the label-free hybridization biosensor could be conveniently used to monitor DNA hybridization with a high sensitivity. X-ray photoelectron spectrometry technique has been employed to characterize the immobilization of single-stranded HBV DNA on a gold surface.