Vitamin D3 up-regulated protein 1(VDUP1) is regulated by FOXO3A and miR-17-5p at the transcriptional and post-transcriptional levels, respectively, in senescent fibroblasts.

Vitamin D(3) up-regulated protein 1 (VDUP1) plays multifunctional roles in diverse cellular responses, particularly in its relation to proliferation, apoptosis, differentiation, and diseases such as cancer and stress-related diseases. In this study, we demonstrated that VDUP1 was up-regulated during the senescence process. Our results showed that overexpression of VDUP1 in young cells caused typical signs of senescence. We also found that VDUP1 knockdown delayed the onset of Ras-induced cellular senescence. Subsequently, we found that FOXO3A, whose activity increased in senescent cells, transcriptionally up-regulates VDUP1 expression and miR-17-5p, whose expression decreased in senescent cells, directly interacted with the 3'-untranslated region of VDUP1 transcripts, and destabilized VDUP1 mRNA in young cells. These results indicated that VDUP1 expression was regulated by FOXO3A at the transcriptional level and by miR-17-5p at the post-transcriptional levels during the senescence process.

[Pathological changes of benign hyperplastic prostate after removal of innervation of cholinergic pelvic nerve: experiment with spontaneous hypertension rats].

OBJECTIVE: To study the pathological change of benign hyperplastic prostate after removal of the innervation of cholinergic parasympathetic pelvic nerve. METHODS: Sixty-five male spontaneous hypertension rats (SHRs) were randomly assigned into 3 groups: operation group (n = 30) undergoing truncation of bilateral originating branches of parasympathetic pelvic nerve of major pelvic ganglion (MPG) followed by cystostomy, sham operation group (operation control group, n = 30) undergoing cystostomy, and normal control group (n = 5) not undergoing operation. 3, 7, 11, 15 and > or = 21 days after operation 6 rats from the 2 operation groups and 1 from the control group were sacrificed to observe the gross morphology and histological and cellular changes of the prostate glands. RESULTS: The prostate of the operation group on post-operational day 7 showed mild granular solidification and such change progressed gradually over time, the ratio of prostate wet weight/rat body weight was (0.4764 +/- 0.0125) mg/g on day 3, then gradually decreased, and became (0.2749 +/- 0.0197) mg/g > or = 21 days post-operationally; while the ratio of prostate tissue dry weight/wet weight on day 3 was (0.1966 +/- 0.0062), then gradually increased, and became (0.2596 +/- 0.0035) > or = 21 days post-operationally. HE staining showed that the glandular structure gradually became dilated and rounded, with accumulation of prostatic fluid. 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. Electron microscopy showed that the glandular cells gradually underwent vacuolar degeneration and the structures of the basement membrane became fuzzy. The smooth muscle cells degenerated mildly, and the fibroblasts and collagenous fibers in the stroma overgrew slowly. All these histological changes were not found in the sham operation control and normal control groups. CONCLUSION: Remarkable atrophy occurs in benign hyperplastic prostatic gland after radical removal of the innervation of cholinergic parasympathetic pelvic nerve. Such operation may represent a novel therapy for BPH.

[Differential transcription of Bcl-2 and Bax through the cell cycle in prostate cancer cell line].

OBJECTIVE: To investigate the differential expression of apoptosis associated gene Bcl-2 and Bax through cell cycle and its possible clinical meaning. METHODS: The prostate cancer cell line PC-3 was synchronized in M, G1, S and G2 phase using modified thymine deoxyriboside blockage and high pressure N2O technique. The efficiency of synchronization was detected by flow-cytometry. RT-PCR and Western blot methods were used to examine the expression of Bcl-2 and Bax in mRNA and protein level. RESULTS: The synchronized rate of M, G1, S and G2 phase were 92.1%, 87.0%, 80.2% and 75.9% respectively. Bcl-2 was constitutively expressed through the cell cycle, but both the mRNA and protein expression level of Bcl-2 were very high in the G1 phase, dramatically decreased in M, S and G2 phase. The expression level of Bax had no change through the cell cycle. CONCLUSIONS: Cell cycle could influence the expression level of Bcl-2 significantly but not Bax, these might have some clinical relevance.

[Expression of ZNRD1 protein in human renal cell carcinoma].

OBJECTIVE: To investigate the expression of zinc ribbon domain-containing1 (ZNRD1) in human renal cell carcinoma and normal kidney tissues. METHODS: The 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. RESULTS: ZNRD1 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). CONCLUSION: ZNRD1 gene and ZNRD1 protein may play an important role in the carcinogenesis of renal cell carcinoma. Further investigation is still needed.

[The pathological change of rats' benign hyperplastic prostate after radical denervation].

OBJECTIVE: To study the pathological change of rats' benign hyperplastic prostate (BHP) after radical denervation. METHODS: A 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. RESULTS: In 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. CONCLUSIONS: After 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.

Rapid identification of UCA1 as a very sensitive and specific unique marker for human bladder carcinoma.

PURPOSE: The most common genitourinary malignancy in China is bladder transitional cell carcinoma (TCC). Early diagnosis of new and recurrent bladder cancers, followed by timely treatment, will help decrease mortality. There are currently no satisfactory markers for bladder cancer available in clinics. Better diagnostic methods are highly demanded. EXPERIMENTAL DESIGN: In this research, we have used comprehensive expressed sequence tag analysis, serial analysis of gene expression, and microarray analysis and quickly discovered a candidate marker, urothelial carcinoma associated 1 (UCA1). The UCA1 gene was characterized and its performance as a urine marker was analyzed by reverse transcription-PCR with urine sediments. A total of 212 individuals were included in this study, 94 having bladder cancers, 33 ureter/pelvic cancers, and 85 normal and other urinary tract disease controls. RESULTS: UCA1 was identified as a novel noncoding RNA gene dramatically up-regulated in TCC and it is the most TCC-specific gene yet identified. The full-length cDNA was 1,439 bp, and sequence analysis showed that it belonged to the human endogenous retrovirus H family. Clinical tests showed that UCA1 assay was highly specific (91.8%, 78 of 85) and very sensitive (80.9%, 76 of 94) in the diagnosis of bladder cancer and was especially valuable for superficial G2-G3 patients (sensitivity 91.1%, 41 of 45). It showed excellent differential diagnostic performance in various urinary tract diseases without TCC. CONCLUSIONS: UCA1 is a very sensitive and specific unique marker for bladder cancer. It could have important implications in postoperative noninvasive follow-up. This research also highlights a shortcut to new cancer diagnostic assays through integration of in silico isolation methods with translational clinical tests based on RNA detection protocols.

[Expression of hypoxia-inducible factor-1-alpha, hypoxia-inducible factor-2alpha and vascular endothelial growth factor in sporadic clear cell renal cell renal cell carcinoma and their significance in the pathogenesis thereof].

OBJECTIVE: To investigate the expression of hypoxia-inducible factor (HIF)-1-alpha, HIF-2alpha, and vascular endothelial growth factor (VEGF) in sporadic clear cell renal cell carcinoma (CCRCC) and to analyze the relationship among them. METHODS: Samples of CCRCC were obtained from 107 patients during resection. Immunohistochemistry was used to detect the expression of HIF-1-alpha, HIF-2alpha, and VEGF in the tumor tissues and normal kidney tissues distant from the tumor. The microvessel density (MVD) was observed by microscopy. RESULTS: HIF-1-alpha and HIF-2alpha were not expressed in the normal kidney tissues. However, in the tumor tissues the HIF-1-alpha positive rate was 69.2% (74/107), significantly lower than that of the HIF-2-alpha (80.5%, 88/107, P = 0.001), the VEGF positive rate was 78.5% (84/107). The VEGF positive rate of the HIF-1-alpha-positive group was 90.5%, significantly higher than that of the HIF-1-alpha-negative group (51.5%, P = 0.001). The VEGF positive rate of the HIF-2-alpha-positive group was 89.8%, significantly higher than that of the HIF-1-alpha-negative group (26.3%, P = 0.001). The MVD of the 64 HIF-1alpha, HIF-2alpha, and VEGF positive samples was 697, significantly higher than that of the 13 HIF-1alpha, HIF-2alpha, and VEGF negative samples (391, P = 0.001). The MVD of the HIF-2alpha positive samples was 678 +/- 324, significantly higher than that of the HIF-2alpha negative samples (383 +/- 293, P = 0.001). The MVD of the VEGF positive samples was 692 +/- 325, significantly higher than that of the VEGF negative samples (384 +/- 269, P = 0.001). Spearman correlation analysis showed that MVD was strongly positively correlated with HIF-2alpha and VEGF. Mann-Whitney U test showed that HIF-1alpha and HIF-2alpha were not correlated with the staging of CCRCC. CONCLUSION: HIF-2alpha is expressed more frequently in sporadic CCRCC than HIF-1alpha. Both HIF-1alpha and HIF-2alpha upregulate the VEGF expression and angiogenesis, especially HIF-2alpha.

[The study of relationship between nuclear metrix protein 22 urinary level and the grade and stage of bladder transitional cell carcinoma].

OBJECTIVE: To assess the relationship between nuclear matrix protein (NMP) 22 urinary level and the grade and stage of bladder transitional cell carcinoma. METHODS: From 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. RESULTS: The 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). CONCLUSIONS: There is a relation between NMP22 urinary level and the grade and stage of bladder transitional cell carcinoma.

[Regulation of expression of pituitary tumor transforming gene 1 (PTTG1) by androgen in prostate cancer].

OBJECTIVE: To identify the androgen-responsive genes in prostate and screen the molecular targets for further studying human prostate cancer. METHODS: The potential androgen-responsive gene pituitary tumor transforming gene 1 (PTTG1) was selected which had been previously screened by cDNA microarray in rat prostate and its mRNA level was detected by Northern blot in the castrated rat prostate with and without replacement of Mibolerone. Immunohistochemistry was performed to determine the expression and location of PTTG1 in human prostate tissues. Then human androgen-dependent prostate cancer cells LNCaP were used as a model to study the regulation of PTTG1 by Mibolerone. RESULTS: PTTG1 mRNA was hardly detectable in the prostate of 7-day castrated rats, while it was up-regulated dramatically in the prostate of 7-day castrated rats treated with Mibolerone for 2 days. It was showed that high expression of PTTG1 was localized to the epithelial cells of human prostate cancer but not to the stromal cells with Immunohistochemistry. Northern blot analysis indicated that LNCaP cells treated with 0.1 nmol/L Mibolerone for 2 days led to the high PTTG1 mRNA expression. The basic expression of PTTG1 in human androgen-independent prostate cancer cell lines PC3 or DU145 was even higher than that in the human androgen-dependent prostate cancer cells LNCaP treated with Mibolerone. CONCLUSION: Androgen can up-regulate the PTTG1 expression in castrated rat prostate and human prostate cancer cell LNCaP. It suggests that PTTG1 is potential to play an important role in human prostate cancer progression.

[Association between A49T polymorphism of SRD5A2 gene and risk of prostate cancer].

OBJECTIVE: To investigate the association between A49T polymorphism of SRD5A2 gene and risk of prostate cancer. METHODS: PCR was used to examine the A49T polymorphisms of SRD5A2 gene in the tissues of prostate cancer resected from 112 patients (CaP group) and the specimens of benign prostate hyperplasia (BPH group) resected from 89 patients. The association of A49T polymorphism with age of onset, FPSA, TPSA, F/T, T stage, and Gleason score were analyzed. RESULTS: There was no significant difference in A49T polymorphism between the CaP and BPH groups (P > 0.05). The average age of CaP patients was significantly higher than that of the BPH patients (P < 0.05). In the CaP patients, the Gleason score was significantly higher, and the age of onset was significantly lower in the AT + TT genotype than in the AA genotype (both P < 0.05) 2. The age of onset of the AA + AT group was significantly lower than that of the AA group (P < 0.05). CONCLUSION: AA + AT genotype may be of worse prognosis, however, without significant difference. Rank scoring may reflect the relation between Gleason score and A49T genotype and estimate the prognosis better than two-level discrete evaluation.

[Association between polymorphism of CYP17 gene and serum hormone concentrations in aged men].

OBJECTIVE: To investigate the association between polymorphism of CYP17 gene and serum hormone concentrations in aged men. METHODS: Eighty-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. RESULTS: No 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). CONCLUSION: A2 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.

[The relationship between acute rejection and expression of sCD30 for the patients after kidney transplantation].

OBJECTIVE: To study the relationship between the sCD30 and acute rejection. METHODS: We tested the sCD30 level in serum for 58 cases with kidney transplantation before and the 7th day and 28th day after operation by ELISA. 31 healthy individual for control group, and simultaneously recorded the incidence of rejection after kidney transplantation. RESULTS: The results showed that there is an obviously relation before kidney transplantation between the sCD30 level in serum and the incidence of acute rejection (chi = 4.843, P = 0.028, P < 0.05). There is a significantly relation at the 7th day after kidney transplantation between the sCD30 level in serum and the incidence of acute rejection (chi = 7.201, P = 0.007, P < 0.01). There is no obviously relation at 28th day after kidney transplantation between the sCD30 level in serum and the incidence of acute rejection (chi = 2.095, P = 0.148, P > 0.05). CONCLUSION: The results suggested that the expressions of sCD30 are related to acute rejection. We speculated that the expressions of sCD30 could play an important role in acute rejection.

Peroxisome proliferator-activated receptor gamma ligands induce cell cycle arrest and apoptosis in human renal carcinoma cell lines.

AIM: To study the effect of peroxisome proliferator-activated receptor gamma (PPARgamma) ligands on cell proliferation and apoptosis in human renal carcinoma cell lines. METHODS: The expression of PPARgamma was investigated by reverse transcriptase polymerase chain reaction (RT-PCR), Western blot and immunohistochemistry. The effect of thiazolidinedione (TZD) PPARgamma ligands on growth of renal cell carcinoma (RCC) cells was measured by MTT assay and flow cytometric analysis. Cell death ELISA, Hoechst 33342 fluorescent staining and DNA ladder assay were used to observe the effects of PPAR gamma ligands on apoptosis. Regulatory proteins of cell cycle and apoptosis were detected by Western blot analysis. RESULTS: PPARgamma was expressed at much higher levels in renal tumors than in the normal kidney (2.16+/-0.85 vs 0.90+/-0.73; P<0.01). TZD PPARgamma ligands inhibited RCC cell growth in a dose-dependent manner with IC50 values of 7.08 micromol/L and 11.32 micromol/L for pioglitazone, and 5.71 micromol/L and 8.38 micromol/L for troglitazone in 786-O and A498 cells, respectively. Cell cycle analysis showed a G0/G1 arrest in human RCC cells following 24-h exposure to TZD. Analysis of cell cycle regulatory proteins revealed that TZD decreased the protein levels of proliferating cell nuclear antigen, pRb, cyclin D1, and Cdk4 but increased the levels of p21 and p27 in a time-dependent manner. Furthermore, high doses of TZD induced massive apoptosis in renal cancer cells, with increased Bax expression and decreased Bcl-2 expression. CONCLUSION: TZD PPAR gamma ligands showed potent inhibitory effect on proliferation, and could induce apoptosis in RCC cells. These results suggest that ligands for PPAR gamma have potential antitumor effects on renal carcinoma cells.

[Expression of SSX2 gene in human urologic neoplasms].

OBJECTIVE: To investigate the expression of SSX(2)gene in human renal cell carcinoma and urinary transitional cell carcinoma. METHODS: Reverse-transcription polymerase chain reaction (RT-PCR) was used for detecting SSX(2) gene in the specimens from renal cell carcinoma (n = 26), urinary transitional cell carcinoma (n = 27) and in 15 specimens taken from the tumor surrounding tissues. RESULTS: Positive expression of SSX(2) gene at mRNA was detected in 69% renal cell carcinomas (18/26), in 81% urinary transitional cell carcinomas (22/27). The mRNA of SSX(2) was not detected in the 15 specimens from tumor surrounding tissues. CONCLUSION: The SSX(2) gene is highly expressed in human renal cell carcinoma and urinary transitional cell carcinoma.

[The expression of hypoxia inducible factor-1,2 alpha in sporadic clear cell renal cell carcinoma and their relationships to the mutations of von Hippel-Lindau gene].

OBJECTIVE: To evaluate the expression of hypoxia inducible factor (HIF)-1alpha, 2alpha in sporadic clear cell renal cell carcinoma and their relationships to the mutations of von Hippel-Lindau (VHL) gene. METHODS: Mutations of VHL gene, expression of HIF-1alpha and 2alpha were detected by polymerase chain reaction (PCR), direct DNA sequencing and immunohistochemistry in 77 cases of Chinese sporadic clear cell renal cell carcinoma (CCRCC). The stage was pT(1)N(0)M(0)in 55 patients (71%), pT(2)N(0)M(0) in 7 patients (9%), pT(3)N(0)M(0) in 14 patients (18%), and pT(4)N(0)M(0) in 1 patient (1%). The classification according to the tumor nuclear grading system showed 15 carcinomas (19%) of tumor nuclear grade 1, 56 (73%) of tumor nuclear grade 2 and 6 (8%) of tumor nuclear grade 3. RESULTS: None of the VHL gene mutations were found in all the normal tissue specimens. VHL gene mutations were detected in 40 (52%) cases of CCRCC. The positive rate of HIF-2alpha (81%) was higher than that of HIF-1alpha (66%) (chi(2) = 23.310, P < 0.01); The positive rate of HIF-1alpha and HIF-2alpha in the cases of mutations (98% and 93% respectively) was higher than that of them in non-mutations (32% and 68% respectively) (chi(2) = 36.386, 7.617, P < 0.01); The correlation between HIF-1alpha and VHL gene mutations was closer than that between HIF-2alpha and VHL gene mutations (partial correlation coefficiency was 4.481 and 2.027 respectively, P < 0.01). The expression of HIF-1alpha and 2alpha in different pathological grade and stage of CCRCC showed no significant difference (P > 0.05). CONCLUSIONS: Our study suggests that VHL gene mutations are frequent in sporadic CCRCC, and the high expression of HIF-1alpha and 2alpha are found in the group of VHL mutations. However, we have not found significant correlation between the expression of HIF-1alpha and 2alpha and pathological grade and stage of CCRCC in our study.

[Short-term neoadjuvant hormone therapy enhanced expression of clusterin in prostate cancer].

OBJECTIVE: To define changes in clusterin expression following short-term neoadjuvant hormone therapy (NHT) and its biological significance in prostate cancer tissues. METHODS: Twenty-six archival radical prostatectomy (RP) specimens without receiving NHT, 19 needle biopsies and corresponding 19 RP specimens following 3-month NHT, were subjected to immunohistochemical clusterin staining. RESULTS: Staining for clusterin was mainly found in cytoplasm and part of extracellular matrix. Clusterin expression was significantly greater in RP specimens with preoperative NHT (t = 2.91, P < 0.01); Needle biopsies obtained before NHT consistently demonstrated lower staining intensity (1.42 +/- 0.51) than corresponding RP specimens (2.16 +/- 0.60) following 3-month NHT (t = 7.10, P < 0.01). CONCLUSIONS: Upregulation of clusterin in part accounts for malignant progression of prostate cancer through its anti-apoptotic action following androgen withdrawal. These findings support that adjuvant therapy targeting clusterin may enhance androgen ablation therapy in advanced prostate cancer.

[Mutation of von Hippel-Lindau gene and expression of vascular endothelial growth factor in sporadic clear cell renal cell carcinoma and their relationships to angiogenesis].

OBJECTIVE: To evaluate the relationship between the mutation of the von Hippel-Lindau (VHL) gene and expression of vascular endothelial growth factor (VEGF) in sporadic clear cell renal cell carcinoma (CCRCC) and angiogenesis. METHODS: Polymerase chain reaction (PCR) was used to detect the mutation of VHL gene in the specimens of cancerous tissue and normal tissues away from tumor from 77 patients with CCRCC. Immunohistochemistry was used to examine the expression of VEGF. CD34 staining was used to measure the microvascular density (MVD). RESULTS: VHL gene mutations were detected in 40 cases (51.9%). The expression rate of VEGF was 79.2% (61 cases). The positive rate of VEGF in the cases with VHL mutation was 92.5%, significantly higher than that in the cases without VHL mutation (64.9%, P = 0.003). The levels of MVD was higher in the cases with VHL mutation and those with VEGF expression were 760.80/mm2 and 715.95/mm2 respectively, both significantly higher than those in the cases without VHL-mutation and those without VEGF expression (547.03/mm2 and 437.44/mm2 respectively, all P = 0.001). The cases with expression of VEGF were divided into two groups according the presence or absence of VHL gene mutations or not. The MVD of the cases with VEGF expression and VHL mutation was 760.80 mm2, significantly higher than that of the cases with VEGF expression and without VHL mutation (547.03 mm2, P = 0.011). CONCLUSION: The mutation rate of VHL gene is high among the Chinese with sporadic CCRCC. VHL gene mutation increases significantly the VEGF expression, thus, and perhaps via other mechanism too, promoting the angiogenesis in tumor. The high level of MVD of the cases with VHL gene mutation may be related to the high malignant potential of CCRCC.

[Prevention of cell death in LNCaP cells by overexpression of clusterin].

OBJECTIVE: To evaluate the effect of clusterin with and without leader sequence on overexpression preventing apoptosis in human prostate LNCaP cells. METHODS: The plasmid pIRES2-EGFP was used to generate the clusterin expression constructs with full-length or without the leader sequence (designated as pIRES2-EGFP/cluac, pIRES2-EGFP/clubc, respectively). Western blot analysis was employed to compare clusterin expression levels in the lysis and supernatant fluid of clusterin transfected LNCaP cells in vitro. The distribution of different functional domains of clusterin in cells was detected with Immunocytochemical staining. The clusterin's protective role of Na2SeO3-induced apoptosis in LNCaP cells was examined by flow cytometry (FCM) and fluorescence microscope. RESULTS: Clusterin expression was detected in the lysis and supernatant fluid of pIRES2-EGFP/cluac transfected LNCaP cells, while clusterin was found only in lysis liquid of pIRES2-EGFP/clubc transfected LNCaP cells, but not found in their supernatant fluid. The distribution of cluserin in the plasm of pIRES2-EGFP/cluac transfected cells was aggregative, and on the other hand, clusterin distributed dispersedly in pIRES2-EGFP/clubc transfected cells. Its anti-apoptotic property in LNCaP cells was proved by FCM and fluorescence microscope. CONCLUSION: It is apparent that clusterin plays an important role in preventing apoptosis in prostate cancer, and the presence of the leader sequence is necessary for clusterin's anti-apoptotic function.

[Frequent somatic mutations of the von Hippel-Lindau (VHL) tumor suppressor gene and its meaning in sporadic human renal clear cell carcinoma].

OBJECTIVE: To investigate the mutation of VHL gene, an important tumor suppressor gene in primary sporadic human renal cell carcinoma (RCC) and analyse its relationships with pathological stage and grade of renal cell carcinoma. METHODS: We analyzed 57 cases of primary sporadic Chinese renal clear carcinoma using the polymerase chain reaction (PCR) and denaturing high performance liquid chromatography(DHPLC). All positive cases in DHPLC analysis were further characterized by direct sequencing. RESULTS: Somatic mutations were detected in 30 (53%) of 57 clear cell renal carcinomas including 13 deletions, 2 insertions, and 15 missense mutations. These mutations mainly occurred in the last one-third region of exon 1, 2,and 3. CONCLUSION: VHL tumor suppressor gene is one of the major tumor suppressor genes in human renal cell carcinoma and there are frequent mutations of VHL in primary sporadic Chinese renal clear cell carcinomas. The mutations of VHL gene were irrespective of the age and pathological grade and stage of patients.

[Peroxisome proliferator-actived receptor-gamma ligand troglitazone induces apoptosis in renal cell carcinoma].

UNLABELLED: OBJECTIVE To investigate the expression of peroxisome proliferator-actived receptor-gamma (PPAR-gamma)and the inducement of apoptosis by PPAR-gamma ligand in renal cell carcinoma(RCC)-derived cell lines. METHODS: RT-PCR and Western blot analysis were performed to determined the expression of PPAR-gamma mRNA and protein in two RCC derived cell lines(786-O and A498) and two normal kidney(NK)-derived cell lines(HK-2 and HMCC). Two RCC cell lines were treated with 50 micromol/L troglitazoned for and evaluated for the effects of antidiabetic thiazolidinediones (TZDs) on the cells apoptosis by fluorescence microscopy and DNA ladder assay. The mutative expressions of Bcl-2 and Bax before and after TZDs treatment were also performed by western blot analysis. RESULTS: The expression of PPAR-gamma was observed to be stronger in 786-O and A498 cells than in HK-2 and HMCC cells by RT- PCR and Western blot analysis. Treated with 50 micromol/L troglitazone (for 48 h) it induced typical apoatosis in 786-O and A498 cells. After treatment, a decrease in Bcl-2 expression in RCC cells was observed by Western blot analysis,and the expression of Bax,however,was up-regulated. CONCLUSION: The results reveal that troglitazone has the tumor-suppressive effect on RCC cells. High-affinity PPAR-gamma ligands (TZDs) may be the candidates for a novel approach to the treatment of this refractory neoplasm.