Objective To evaluate the expression and significance of differential expression gene Hepsin in the prostate cancer (PC) screened by the cDNA microarray technique.Methods The techniques of semiquantitative RT-PCR and Western blotting were used to detect the mRNA and protein expression of Hepsin. Specimens of 40 cases of PC, 15 benign prostatic hyperplasia (BPH) and 6 normal prostate tissues were examined by the immunohistochemical stain.Results Hepsin was more expressed in PC tissue than normal prostate tissue (P=0.026) and was confirmed by Western blot analysis. Immunohistochemical test confirmed this and demonstrated that Hepsin did not expressed in normal prostate but expressed in PC and BPH and there was a significant difference in Hepsin expression level between PC and BPH tissues ( P=0.000).Conclusion Hepsin high expressed in PC may be a new molecular marker in early diagnosis of PC and a new target for gene therapy of PC.

OBJECTIVETo investigate the expression of zinc ribbon domain-containing1 (ZNRD1) in human renal cell carcinoma and normal kidney tissues.

METHODSThe expression of ZNRD1 protein was examined by immunohistochemical staining in 71 renal cell carcinomas and 24 samples of normal kidney tissue. The correlation between the expressions of ZNRD1 protein and clinicopathologic features was analyzed. The expression of ZNRD1 mRNA and ZNRD1 protein was detected by quantitative reverse transcriptase-polymerase chain reaction (PT-PCR) and Western blot in 20 renal cell carcinomas and corresponding adjacent non-cancerous tissues.

RESULTSZNRD1 protein was detected mostly in the cell nuclei by immunohistochemistry. The positive expression rate of ZNRD1 protein was 91.7% (22/24) in renal cell carcinomas and 20.8% (5/24) in the normal kidney tissues, with a statistically significant difference between cancer and normal kidney tissue (P < 0.01). However, no significant correlation was observed between ZNRD1 protein expression level and clinicopathologic features (P > 0.05). ZNRD1 mRNA expression level was significantly higher in renal cell carcinomas (0.6186) than that in the normal kidney tissues (0.4273) assessed by RT-PCR (P < 0.01). The expression level of ZNRD1 protein by Western blot was 0.5623 in renal cell carcinomas, significantly higher than that in normal kidney tissues (0.3885, P < 0.01).

CONCLUSIONZNRD1 gene and ZNRD1 protein may play an important role in the carcinogenesis of renal cell carcinoma. Further investigation is still needed.

Adult ; Aged ; Aged, 80 and over ; Blotting, Western ; Carcinoma, Renal Cell ; metabolism ; pathology ; DNA-Binding Proteins ; biosynthesis ; genetics ; Female ; Humans ; Immunohistochemistry ; Kidney ; metabolism ; Kidney Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; Neoplasm Staging ; RNA, Messenger ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

Objective To investigate the anti-apoptotic function of clusterin in LNCaP cell line and the role of clusterin antisense oligodcoxynucleotide(AS-ODN)in TNF-?-induced death of LNCaP cell. Methods The wild type LNCaP(group L),LNCaP transfected with the control vector(group M),LNCaP transfected with full-length clusterin expression vector(group A,ie,study group)were cultured.For the de- tection of cytotoxic effect of TNF-?,MTT and ELISA methods were used to determine the cell proliferation and apoptosis of the 3 clones,and the changes of proliferation and apoptosis in A cell after transfection of clusterin AS-ODN were also assessed.Results MTT method showed that the cell proliferation activity(A value)of groups L,M,and A were 0.84?0.03,0.85?0.04,0.95?0.03,respectively;the difference be- tween groups L and M was not significant(P＞0.05);but compared with group A the cell proliferation activ- ity was significantly lower in groups L and M(P＜0.01 for both).ELISA resuhs showed that the A values of groups L,M,and A were 0.59?0.04,0.62?0.03,0.33?0.04,respectively;the difference between groups L and M was not significant(P＞0.05);but compared with group A,the apoptosis rates were significantly higher in groups L and M(P＜0.01 for both).In group A,A values of cell proliferation activity in subgroups control,AS-ODN,TNF-?,TNF-?+AS-ODN were 1.30?0.03,1.25?0.03,0.99?0.03,0.80?0.03, respectively;the differences between each group were significant(P＜0.05 for all).And the A values of cell apoptosis in the above 4 groups were 0.02?0.00,0.21?0.02,0.63?0.07,1.16?0.04,respectively,the differences between each group were significant(P＜0.01 for all).Conclusions Stable transfection and subsequent expression of clusterin result in resistance to the cytotoxic effect of TNF-?.Transfection with clus- terin AS-ODN enhances cytotoxic effect of TNF-?in A cells.These results suggest that clusterin plays an im- portant role in anti-apoptotic function in LNCaP cell line.

Carnosic acid (CA) is the main phenolic diterpenoid active ingredient in plants such as rosemary and sage, and has antiviral, antioxidant, anti-inflammatory effects and so on, however, its antiviral activity against influenza virus infections was not reported. In this study, antiviral activities against influenza A virus infections of three main bioactive ingredients from rosemary, including rosmarinic acid, CA and ursolic acid, were evaluated using virus titer titration assay, and CA showed remarkable inhibition on influenza H5N1 replication in A549 cells. The antiviral activity of CA was further confirmed and its mechanism of action was investigated using the indirect immunofluorescence assay (IFA), Western blot and real-time fluorescence quantification polymerase chain reaction (qRT-PCR). The results showed that the 50% effective concentration (EC₅₀) of CA against influenza H5N1 in A549 cells and MDCK cells were 4.30 and 3.64 μmol·L^-1, respectively. Meanwhile, CA also showed inhibition on influenza virus 2009panH1N1 (EC₅₀: 10.1 μmol·L^-1) and H3N2 (EC₅₀: 12.8 μmol·L^-1) replications in A549 cells. Mechanistic studies showed that antiviral activity of CA is related to its induction of heme oxygenase-1 (HO-1) in A549 cells and suppression on production of reactive oxygen in H5N1-infected cells.

Objective To investigate the effects of dominant-negative truncation mutant?NTCF4, lacking the N-terminal form of TCF4 gene,on biological characteristics of renal cancer cell line GRC-I and explore the molecular mechanisms.Methods GRC-I cell was transfected with pCDNA3-?NTCF4 eukary- otie expression plasmid,pCDNA3 empty vector to construct the stable cell line GRC-I/?NTCF4 and GRC-I/ Mock respectively.The morphological changes of stable cells were observed and the cells growth curve was detected through light microscope.The cellular proliferation activities were determined using the MTT assay. The protein expression of Wnt pathway downstream target gene C-Myc and Cox-2 was evaluated by immuno- cytoehemieal method and Western Blot analysis.Results After the dominant-negative?NTCF4 gene was permanently expressed,the GRC-I/?NTCF4 stable cells morphologically showed that appearance changed from circular to long-spindle shape,growth rate decreased with less karyosehisis found,malignant pheno- types reversed to normal renal tubular cells.MTT assay revealed that the proliferation activities of GRC-1/?NTCF4 cells were inhibited by 11.2%-35.5% compared with GRC-I cells (P＜0.05),while the GRC- I/Mock cells have no difference with the control cells.Immunocytochemical analysis and Western Blot showed that the C-Myc and Cox-2 protein expression level of GRC-I/?ANTCF4 cells were significantly sup- pressed in comparison with that of GRC-I/Mock and GRC-I cells.Conclusions The dominant-negative truncation mutant?NTCF4 could partially inhibit the growth of renal cancer cells and down-regulate the pro- tein expression of Wnt pathway target gene C-Myc and Cox-2.These findings provide a experimental founda- tion for applying cell signal therapy to renal cell cancer by blocking the Wnt signaling pathway.

OBJECTIVETo study the paracrine effect of the neuroendocrine differentiation cells and its influence on androgen receptor expression of prostate carcinoma cells.

METHODSEstablished an in vitro induced model of neuroendocrine differentiation of prostate carcinoma (LNCaP(NE), PC-3M(NE)), and investigate the proliferation effect of neuroendocrine phenotype cells (LNCaP(NE), PC-3M(NE)) on other non-neuroendocrine phenotype cells (LNCaP, PC-3M) by feeding non-neuroendocrine phenotype cells with the medium pre-incubated with induced neuroendocrine phenotype cells. It was also tested the regulation of androgen receptor mRNA and androgen receptor protein expression of LNCaP(NE) cells on LNCaP cells by reverse transcriptase polymerase chain reaction and Western Blot in the presence or absence of androgens.

RESULTSThe medium pre-incubated with PC-3M(NE) cells could promote the proliferation of PC-3M cells. The medium pre-incubated with LNCaP(NE) cells could promote the proliferation of LNCaP cells, and reduce the expression of androgen receptor in the latter in the absence of androgens, but the negative results were observed in the presence of androgens.

CONCLUSIONSThe neuroendocrine phenotype cells of prostate cancer can reduce the expression of androgen receptor in prostate cancer cells and promote them to proliferate by means of paracrine in the blockade of androgens.

Androgens ; pharmacology ; Cell Division ; drug effects ; Cell Line, Tumor ; Humans ; Male ; Neoplasms, Hormone-Dependent ; metabolism ; pathology ; Prostatic Neoplasms ; metabolism ; pathology ; Receptors, Androgen ; metabolism

OBJECTIVETo study the pathological change of rats' benign hyperplastic prostate (BHP) after radical denervation.

METHODSA total of 65 male spontaneous hypertension rats (SHR) at 30 weeks age were randomly assigned into treatment group, sham surgery control group and normal control group. In surgery group, all the axonal branches of the major pelvic ganglion (MPG) supplying the bilateral prostate were truncated, followed performing of cystostomy; In sham surgery control group, only cystostomy was performed; In normal control group, no procedure was performed. The rats were sacrificed at 3, 7, 11, 15 and >or= 21 d post-operation respectively. The gross morphological changes of prostate in all animals were observed.

RESULTSIn treatment group, the prostate in 3 d post-operation showed granular solidification and shrunken volume and the changes occurred gradually over time. The glandular epithelial cells showed gradual degeneration, necrosis and detachment. The glandular epithelium became progressively thinner, the smooth muscles elongated and thinned progressively and the stromal components showed mild to moderate overgrowth. At the later stage, the glandular epithelium, glandular lumen and smooth muscles gradually disappeared and the prostate was largely replaced by connective tissues. Electron microscopic study showed that the glandular cells gradually underwent vacuolar degeneration and the structures of basement membrane became fuzzy. The smooth muscles cells degenerated overtime and the fibroblasts and collagenous fibers in the stroma overgrew slowly. At the late stage, most of the glandular cells became necrotic, the basal membrane and smooth muscle cells disappeared and collagenous fibers were highly hyperplasic. In surgery group in 3 d post-operation, the S-100 staining of nerve fiber was diffuse and disappeared after 11 d while it persisted normally in other groups. The two values in sham surgery control group showed no significant changes post-operatively.

CONCLUSIONSAfter radical denervation, the rat prostate with benign hyperplasia (gland and smooth muscles) undergoes dramatic atrophic changes and the volume decreases significantly. It suggests that this treatment may represent a novel therapy for BPH.

Animals ; Denervation ; methods ; Disease Models, Animal ; Male ; Microscopy, Electron, Transmission ; Prostate ; innervation ; pathology ; ultrastructure ; Prostatic Hyperplasia ; pathology ; surgery ; Random Allocation ; Rats ; Rats, Inbred SHR

OBJECTIVETo assess the relationship between nuclear matrix protein (NMP) 22 urinary level and the grade and stage of bladder transitional cell carcinoma.

METHODSFrom June 1999 to March 2005 642 patients underwent NMP22 test, and then the test by cystoscope and pathology were performed in 1 week to 1 month. According to the pathological grade, the patients were divided into 3 groups: group G(1): 69 cases, male 58 and female 11; group G(2): 375 cases, male 255 and female 120; group G(3): 198 cases, male 143 and female 55. And the difference of NMP22 between each group were compared. Meanwhile, according to pathological stage, 239 patients were divided into 3 groups: group PT(1): 121 cases, male 76 and female 45; group PT(2): 65 cases, male 37 and female 28; group PT(3): 53 cases, male 29 and female 24. And the difference of NMP22 between each group were compared.

RESULTSThe concentrations of NMP22 had significant difference between the 3 groups which divided according to pathological grade (Kruskal-Wallis test chi(2) = 67.547, P < 0.001); The concentrations of NMP22 had significant difference between the 3 groups which divided according to pathological stage (Kruskal-Wallis test chi(2) = 20.629, P < 0.001).

CONCLUSIONSThere is a relation between NMP22 urinary level and the grade and stage of bladder transitional cell carcinoma.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Biomarkers, Tumor ; urine ; Carcinoma, Transitional Cell ; pathology ; urine ; Female ; Humans ; Male ; Middle Aged ; Neoplasm Staging ; Nuclear Proteins ; urine ; Urinary Bladder Neoplasms ; pathology ; urine

OBJECTIVESTo investigate Bsm I single nucleotide polymorphism (SNP) of vitamin D receptor gene (VDRG) in low-risk Chinese Han population and its relationship to the susceptibility to prostate cancer (PCa), and to discuss the possible reason for the racial difference of PCa.

METHODSOne hundred and three patients with PCa and 106 normal controls, mainly from Northern Chinese Han population, were enrolled in this study. Their blood samples were obtained, all of which were genotyped for Bsm I SNP by denaturing high performance liquid chromatography(DHPLC) methods using case-control study.

RESULTSThe distribution of genotype and allele had no significant difference between PCa patients and normal controls (P > 0.05). The frequencies for the bb, Bb and BB genotypes in PCa patients and normal controls were 92.23%/94.34%, 7.77%/5.66%, and 0/0, respectively. The frequencies for B and b allele were 3.88%, 96.12% and 2.91%, 97.09%, respectively.

CONCLUSIONSThe results indicate no significant relationship between the VDRG polymorphisms and PCa in Northern Chinese Han population. The distribution of VDRG Bsm I SNP varies in different ethnic populations, which may be one reason for the racial difference of PCa.

Aged ; Aged, 80 and over ; Case-Control Studies ; China ; ethnology ; Chromatography, High Pressure Liquid ; Genetic Predisposition to Disease ; Genotype ; Humans ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; Prostatic Neoplasms ; genetics ; Receptors, Calcitriol ; genetics

OBJECTIVETo investigate the association between polymorphism of CYP17 gene and serum hormone concentrations in aged men.

METHODSEighty-three healthy men at the average age of 66.7 were divided into a < 66.7 group (n = 36) and a > 66.7 group (n = 47), and the polymorphism of CYP17 gene in the 5' promoter region was investigated by PCR using DNA from the men's peripheral blood lymphocytes. A new recognition site was created for the restriction enzyme MspA1 I by transition (T --> C) in the risk allele (A2). Three genotypes A1/A1, A1/A2, A2/A2 were established, serum sex-hormone levels measured, and mean hormone concentration evaluated in each genotype and age group.

RESULTSNo evidence was found that the testosterone (T) level, estrogen (E2) level and T/E2 ratio were associated with the genotype of CYP17 gene. There was no significant difference in T and E2 levels between the two groups, but there was a significant increase in the T/E2 ratio (P < 0.05).

CONCLUSIONA2 allele does not increase sex hormone levels in aged men, but the T/E, ratio was higher in the > 66.7 group than in the < 66.7 group. This may be closely associated with the mechanism of benign prostate hyperplasia and prostate cancer in aged men.

Adult ; Aged ; Aged, 80 and over ; Estradiol ; blood ; Genotype ; Humans ; Male ; Middle Aged ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Promoter Regions, Genetic ; genetics ; Steroid 17-alpha-Hydroxylase ; genetics ; Testosterone ; blood