Exosomal PGAM1 promotes prostate cancer angiogenesis and metastasis by interacting with ACTG1.

Tumor-derived exosomes and their contents promote cancer metastasis. Phosphoglycerate mutase 1 (PGAM1) is involved in various cancer-related processes. Nevertheless, the underlying mechanism of exosomal PGAM1 in prostate cancer (PCa) metastasis remains unclear. In this study, we performed in vitro and in vivo to determine the functions of exosomal PGAM1 in the angiogenesis of patients with metastatic PCa. We performed Glutathione-S-transferase pulldown, co-immunoprecipitation, western blotting and gelatin degradation assays to determine the pathway mediating the effect of exosomal PGAM1 in PCa. Our results revealed a significant increase in exosomal PGAM1 levels in the plasma of patients with metastatic PCa compared to patients with non-metastatic PCa. Furthermore, PGAM1 was a key factor initiating PCa cell metastasis by promoting invadopodia formation and could be conveyed by exosomes from PCa cells to human umbilical vein endothelial cells (HUVECs). In addition, exosomal PGAM1 could bind to γ-actin (ACTG1), which promotes podosome formation and neovascular sprouting in HUVECs. In vivo results revealed exosomal PGAM1 enhanced lung metastasis in nude mice injected with PCa cells via the tail vein. In summary, exosomal PGAM1 promotes angiogenesis and could be used as a liquid biopsy marker for PCa metastasis.

LncRNA SNHG1 and RNA binding protein hnRNPL form a complex and coregulate CDH1 to boost the growth and metastasis of prostate cancer.

The interaction between LncRNA and RNA-binding protein (RBPs) plays an essential role in the regulation over the malignant progression of tumors. Previous studies on the mechanism of SNHG1, an emerging lncRNA, have primarily focused on the competing endogenous RNA (ceRNA) mechanism. Nevertheless, the underlying mechanism between SNHG1 and RBPs in tumors remains to be explored, especially in prostate cancer (PCa). SNHG1 expression profiles in PCa were determined through the analysis of TCGA data and tissue microarray at the RNA level. Gain- and loss-of-function experiments were performed to investigate the biological role of SNHG1 in PCa initiation and progression. RNA-seq, immunoblotting, RNA pull-down and RNA immunoprecipitation analyses were utilized to clarify potential pathways with which SNHG1 might be involved. Finally, rescue experiments were carried out to further confirm this mechanism. We found that SNHG1 was dominantly expressed in the nuclei of PCa cells and significantly upregulated in PCa patients. The higher expression level of SNHG1 was dramatically correlated with tumor metastasis and patient survival. Functionally, overexpression of SNHG1 in PCa cells induced epithelial-mesenchymal transition (EMT), accompanied by down-regulation of the epithelial marker, E-cadherin, and up-regulation of the mesenchymal marker, vimentin. Increased proliferation and migration, as well as accelerated xenograft tumor growth, were observed in SNHG1-overexpressing PCa cells, while opposite effects were achieved in SNHG1-silenced cells. Mechanistically, SNHG1 competitively interacted with hnRNPL to impair the translation of protein E-cadherin, thus activating the effect of SNHG1 on the EMT pathway, eventually promoting the metastasis of PCa. Our findings demonstrate that SNHG1 is a positive regulator of EMT activation through the SNHG1-hnRNPL-CDH1 axis. SNHG1 may serve as a novel potential therapeutic target for PCa.

[Expression of phosphoribosyl pyrophosphate synthase 2 in human testis tissue].

OBJECTIVE: To investigate the expression of phosphoribosyl pyrophosphate synthase 2 (PRPS2) in the human testis and its clinical significance. METHODS: Using quantitative real-time PCR (qRT-PCR) and immunohistochemistry, we detected the expression of PRPS2 mRNA in the testis tissue of the men with normal spermatogenesis or mile, moderate or severe hypospermatogenesis (HS) and that of the PRPS2 protein in the testicular biopsy tissue of 67 adult males. Then, we analyzed the relationship of the PRPS2 expressions with the testicular histological types and clinical parameters of the subjects. RESULTS: The expression of PRPS2 mRNA in the testis tissue was significantly higher in the normal spermatogenesis group than in the moderate and severe HS groups (P < 0.01). The positive expression of the PRPS2 protein was 70.0% in the normal spermatogenesis group, 66.7% in the mild HS group, 50.0% in the moderate HS group and 23.8% in the severe HS group, significantly higher in the normal spermatogenesis and mild HS groups than in the moderate and severe HS groups (P < 0.01). No significant correlation, however, was observed between the PRPS2 expression and clinical parameters of the subjects (P > 0.05). CONCLUSIONS: PRPS2 is lowly expressed in the testis tissue of the men with hypospermatogenesis and its expression level may help the diagnosis of male infertility and the prediction of the spermatogenic function of the testis.

Phosphoribosyl pyrophosphate synthetases 2 knockdown inhibits prostate cancer progression by suppressing cell cycle and inducing cell apoptosis.

Phosphoribosyl pyrophosphate synthetases 2 (PRPS2) protein function as nucleotide synthesis enzyme that plays vital roles in cancer biology. However, the expression profile and function of PRPS2 in prostate cancer (PCa) remain to be identified. Here we investigated the expression of PRPS2 protein in human PCa and paired normal tissues by immunohistochemistry, meanwhile the regulatory effects on cell proliferation, apoptosis and growth of xenograft tumors in nude mice were evaluated in PCa cells with PRPS2 depletion. Moreover, the signaling pathways were also explored by western blot analysis and quantitative polymerase chain reaction assays. We found that PRPS2 was dramatically upregulated in prostate adenocarcinoma tissues in comparison with normal tissues, and that increased PRPS2 was linked intimately to advanced clinical stage and pT status. Functional experiments showed that knockdown of PRPS2 significantly suppressed cell growth both in vitro and in vivo. In addition, depletion of PRPS2 induced G1 phase cell cycle arrest and elevated cell apoptosis. Silencing of PRPS2 resulted in the decreased expression of Bcl­2 and cyclinD1 and increased levels of Bax, cleavage of caspases­3, caspases­9 and PARP. Furthermore, we also detected PRPS2 expression was significantly induced after DHT treatment, which implied the important role of PRPS2 in oncogenesis of PCa. Taken together, our findings elucidated that PRPS2 may be a potential novel candidate for PCa therapy.

Phosphoribosyl-pyrophosphate synthetase 2 (PRPS2) depletion regulates spermatogenic cell apoptosis and is correlated with hypospermatogenesis.

Phosphoribosyl-pyrophosphate synthetase 2 (PRPS2) is a rate-limiting enzyme and plays an important role in purine and pyrimidine nucleotide synthesis. Recent studies report that PRPS2 is involved in male infertility. However, the role of PRPS2 in hypospermatogenesis is unknown. In this study, the relationship of PRPS2 with hypospermatogenesis and spermatogenic cell apoptosis was investigated. The results showed that PRPS2 depletion increased the number of apoptotic spermatogenic cells in vitro. PRPS2 was downregulated in a mouse model of hypospermatogenesis. When PRPS2 expression was knocked down in mouse testes, hypospermatogenesis and accelerated apoptosis of spermatogenic cells were noted. E2F transcription factor 1 (E2F1) was confirmed as the target gene of PRPS2 and played a key role in cell apoptosis by regulating the P53/Bcl-xl/Bcl-2/Caspase 6/Caspase 9 apoptosis pathway. Therefore, these data indicate that PRPS2 depletion contributes to the apoptosis of spermatogenic cells and is associated with hypospermatogenesis, which may be helpful for the diagnosis of male infertility.

[Video endosopic inguinal lymphadenectomy for penile cancer].

Inguinal lymph node metastasis is one of the important factors affecting the prognosis of penile cancer. Conventional open inguinal lymphadenectomy, with a high rate of complications, seriously affects the effect of surgery and the patient's quality of life, and therefore is rarely employed nowadays as a treatment option. Video endosopic inguinal lymphadenectomy (VEIL), however, can significantly reduce the incidence rate of surgery-related complications, achieve a desirable control of the tumor, and markedly improve the prognosis. This review focuses on the application, development, indications, effectiveness and complications of VEIL in the treatment of penile cancer.

Downregulation of lncRNA PVT1 expression inhibits proliferation and migration by regulating p38 expression in prostate cancer.

Long non-coding RNA (lncRNA) plasmacytoma variant translocation 1 (PVT1) has been reported to be overexpressed in prostate cancer cells and associated with tumorigenesis in various types of cancer. However, the biological function of lncRNA PVT1 remains largely unknown. The aim of the present study was to investigate the effect of lncRNA PVT1 expression on the proliferation and migration of prostate cancer cells. Stably transfected prostate cancer cells with downregulated expression of lncRNA PVT1 were constructed by an efficient siRNA fragment, followed by confirmation by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Proliferation was assessed using CCK-8, colony formation and xenograft assays, and cell migration was evaluated using a wound healing assay. The PathScan® Intracellular Signaling Array kit was utilized to explore the underlying molecular mechanisms of lncRNA PVT1 expression in prostate cancer cells. RT-qPCR results confirmed that the lncRNA PVT1 expression level was successfully knocked down in prostate cancer cells. When lncRNA PVT1 expression was downregulated in prostate cancer cells, proliferation and migration were significantly inhibited, compared with the control lncRNA PVT1 group. Furthermore, PVT1 knockdown decreased the phosphorylation of p38 in DU145 cells. Therefore, the present study demonstrated that lncRNA PVT1 downregulation inhibits the proliferation and migration of prostate cancer cells, and is associated with p38 phosphorylation.

Pure small-cell carcinoma of the prostate presenting with increasing prostate-specific antigen levels: A case report and review of the literature.

The incidence of prostatic cancer (PCa) has increased significantly, and the measurement of prostate-specific antigen (PSA) is an effective screening tool for its diagnosis. PCa includes a number of specific clinicopathological types, including squamous cell, urothelial, adenoid cystic and small-cell carcinoma, among which small-cell carcinoma of the prostate (SCCP) is extremely rare, accounting for <0.5% of all PCa cases. SCCP is very aggressive and the majority of the cases have a poor prognosis, with a mean survival of ~5 months; it also exhibits specific clinicopathological characteristics and may be divided into two subtypes, namely pure and mixed SCCP. According to the previous literature on PubMed, pure SCCP is not associated with an increase in serum PSA levels. However, the rare case presented herein exhibited an increasingly abnormal serum PSA level. The patient was aged 66 years and had a PSA level of 56.78 ng/ml (normal, <4 ng/ml); he was diagnosed with pure SCCP, underwent radical prostatectomy and has remained disease-free during the follow-up. Similar cases previously published on PubMed were also reviewed, and considerations of survival status and treatment options were analyzed.

[Influences of three surgical approaches to urethral stricture on the erectile function of the patients].

OBJECTIVE: To evaluate the impacts of three different surgical approaches to urethral stricture on the erectile function of the patients. METHODS: This study included 126 male patients with urethral stricture, 35 treated by substitution urethroplasty (group A), 52 by anastomotic urethroplasty (group B), and 39 by internal urethroplasty (group C). We evaluated the pre- and postoperative erectile function of the patients using IIEF-5 scores by telephone calls and interviews. We also monitored their nocturnal penile tumescence (NPT). RESULTS: The IIEF-5 scores in groups A, B and C were 13.5 +/- 4.5, 11.1 +/- 4.8 and 14.5 +/- 4.41 respectively after surgery, all significantly decreased as compared with 17.1 +/- 2.6, 17.1 +/- 3.0 and 17.6 +/- 2.2 preoperatively (P < 0.05). CONCLUSION: All the three surgical approaches can reduce IIEF-5 scores in patients with urethral stricture, but anastomotic urethroplasty may induce a higher incidence of erectile dysfunction than the other two approaches.

[Pathogeneses of erectile dysfunction after rectal cancer treatment].

Rectal cancer is a common malignancy in the alimentary tract with an increasing incidence, the current treatments of which include surgery, radiotherapy, chemotherapy, and integrated comprehensive options. Sexual dysfunction, especially erectile dysfunction (ED), is one of the commonest complications in men after rectal cancer treatment and is generally attributed to the damage to the pelvic autonomic nerves. However, recent studies show that ED after rectal cancer treatment is a complex pathophysiological process associated with neurogenic, vasculogenic, and psychological factors. This article reviews the pathogeneses of ED after rectal cancer treatment in order to provide some theoretical evidence for its prevention and treatment.

[The highly expressed secreted phosphoprotein 1 gene in prostate cancer metastasis: a microarray-based bioinformatic analysis].

OBJECTIVE: To investigate the composition, function, and regulatory mechanisms of the secreted phosphoprotein 1 (SPP1) gene in metastatic prostate cancer. METHODS: We obtained the data about the whole genomic expression profiles on prostate cancer metastasis from the GEO database, and performed data-mining and bioinformatic analysis using BRB-Array Tools and such softwares as Protparam, MotifScan, SignalP 4.0, TMHMM, NetPhos2.0, PredictProtein, GO, KEGG, and STRING. RESULTS: Totally, 73 co-expressed differential genes in prostate cancer metastasis were identified, 21 up-regulated and 52 down-regulated (P <0.01). Bioinformatic analysis indicated that the highly expressed SPP1 gene encoded 314 amino acids and contained 2 N-glycosylation sites, 8 casein kinase II phosphorylation sites and 3 protein kinase C phosphorylation sites, playing essential roles in extracellular matrix (ECM) binding, ossification, osteoblast differentiation, cell adhesion, PI3K-Akt signaling pathway, focal adhesion, Toll-like receptor signaling pathway, and ECM-receptor interaction. CONCLUSION: The bioinformatic method showed a high efficiency in analyzing microarray data and revealing internal biological information. SPP1 may play an important role in prostate cancer metastasis and become a novel biomarker for the diagnosis of prostate cancer metastasis and a new target for its treatment.

[Changes in prostatic stromal composition and benign prostatic hyperplasia].

OBJECTIVE: To investigate whether there are different stromal compositions in the prostate tissue of patients with benign prostatic hyperplasia (BPH) and evaluate their significance in the course of the disease. METHODS: Forty-three surgical or bioptic prostatic specimens of BPH and 5 autoptic normal prostatic specimens were stained by the Masson method to display the elements of the muscle fiber and collagen. The relationship of the changes in the prostatic stromal composition was analyzed with the degree of bladder outlet obstruction (BOO) , IPSS and medication results. RESULTS: The mean ratio of muscle fiber to collagen in the normal prostate tissue was (3.2 +/- 0.2):1, significantly higher than that of the BPH patients (1: [4.7 +/- 3.1] ) (P < 0.01); that in the BPH patients with BOO was 1: (5.4 +/- 3.7) markedly lower than in those without BOO (1: [2.5 +/- 1.1] ) (P = 0.02); that in the BPH patients with severe prostatic symptoms was 1: (9.1 +/- 2.9), remarkably lower than in those with moderate (1: [5.3 +/- 3.4]) and mild prostatic symptoms (1: [2.8 +/- 1.7]) (P < 0.01); and that in the BPH patients with satisfactory medicinal therapeutic results was 1:(2.3 +/- 1.9), significantly higher than in those with poor therapeutic results (1: [7.6 +/- 4.3]) (P < 0.01). CONCLUSION: The stromal composition in the prostatic tissue of BPH patients undergoes different degrees of changes. More obvious BPH symptoms and poorer therapeutic results are associated with a bigger proportion of collagens and a smaller proportion of muscle fibers in the prostatic tissue. These changes may play an important role in the development and progression of BPH.

Patient outcome and prognostic factors of renal cell carcinoma in clinical stage T(1-3)N(1-2)M(0): a single-institution analysis.

OBJECTIVE: To report our data of patients with clinical stage T(1-3)N(1-2)M(0) renal cell carcinoma (RCC) and explore the biological behavior of this malignancy. METHODS: A total of 531 patients with no distant metastatic RCC underwent open radical nephrectomy at our institution between 1988 and 2008, among whom 42 patients with histological nodal metastases had successful surgical tumor resection. The clinical data and outcomes of the 42 patients were analyzed. RESULTS: Of those 42 patients, 19.0% had T1, 21.4% had T2, and 59.5% had T3 stage tumors; 42.9% had N1 and 57.1% had N2 stage tumors. Tumor recurred in 30 (71.4%) patients after the surgery, and death occurred in 26 (61.9%) cases at the last follow-up; among the recurrent cases, 83.3% (25/30) had multiple metastases at the initial recurrence. The median cancer-specific survival (CSS) and disease-free survival (DFS) was 23 and 11 months in these cases, respectively. Multivariate analysis demonstrated that Fuhrman grade (P=0.005), N stage (P=0.014) and T stage (P=0.037) were the independent predictors of CSS; Eastern Cooperative Oncology Group (ECOG) performance status (PS) (P=0.002), tumor size (P=0.007), Fuhrman grade (P=0.009) and N stage (P=0.019) were the independent predictors of DFS. CONCLUSION: Patients with T(1-3)N(1-2)M(0) RCC have poor prognosis. N stage is an independent predictor of both CSS and DFS, suggesting that extended lymph node dissection should be performed when suspicious enlarged nodal disease is found during surgery.

[Expressions of cysteine-rich secretory protein 2 in asthenospermia].

OBJECTIVE: To investigate the mRNA and protein expression levels of cysteine-rich secretory protein 2 (CRISP2) in the sperm of asthenospermia patients, and explore their relationship with sperm motility and related molecular mechanism. METHODS: We collected 78 semen samples from adult male patients with asthenospermia and another 70 from healthy volunteers as controls. We extracted total RNA and total protein from the sperm following purification of the sperm by Percoll gradient centrifugation, and detected the relative expressions of CRISP2 mRNA and protein in the two groups by RT-PCR, SYBR Green real-time PCR and Western blot. RESULTS: The expression of CRISP2 mRNA was down-regulated by 4.3 times and that of the CRISP2 protein by 1.71 times in the asthenospermia patients, significantly lower than in the normal control group (P < 0.05). CONCLUSION: The down-regulation of CRISP2 mRNA and protein expressions in the sperm of asthenospermia patients may be closely related with decreased sperm motility, which suggests that CRISP2 may serve as a potential molecular target for the research of asthenospermia.

[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.

[Single- and two-layer gradient centrifugation in sperm separation: comparison and appraisal].

OBJECTIVE: To appraise the effect of single- and two-layer Percoll density gradient centrifugation in sperm separation. METHODS: Twenty semen specimens underwent single-(50%) and two-layer (90% and 45%) density gradient centrifugation, respectively. The sperm class analyzer (SCA) was used to analyze sperm density, motility and dynamic parameters and round cell density before and after the treatment. RESULTS: After separation, the sperm recovery rate of the single-layer method was (65.5 +/- 12.8)%, significantly higher than that of the two-layer method (P < 0.01). The percentages of grade a sperm of the single- and two-layer method were significantly higher than pre-treatment (P < 0.05, P < 0.01), that of the single-layer was significantly lower than that of the two-layer method (P < 0.05), but the percentage of grade c sperm of the former was significantly higher than that of the latter (P < 0.05). Compared with pre-treatment, the percentage of grade a + b sperm of the two-layer method was significantly higher (P < 0.05), while that of the single-layer method showed no significant difference (P > 0.05), and the round cell density of both the methods was significantly lower (P < 0.05, P < 0.01), with no significant differences between the two methods (P > 0.05). CONCLUSION: The single-layer method yields a higher rate of sperm recovery and causes little change in the sperm motility, while the two-layer method effects a lower rate and significantly improves sperm motility. Both the methods can efficiently separate sperm from round cells, and each has its own advantages and its application value in in vitro treatment of sperm.

[Update of asthenospermia-related genes and proteins].

One of the most common causes of male infertility is asthenospermia, whose pathogenesis, however, is not yet clear. Recent researches have found that some genes (such as tektin-2, DNAI1, DNAH5, DNAH11, AKAP4, SEPT4 and Smcp) and proteins (such as sperm proteins ACTB, ANXA5, PRM1, PRM2 and SABP and seminal proteins Tf, PSA, PAP and Fractalkine) are associated with asthenospermia. The finding of these molecular markers has provided a base for the explanation of the molecular mechanism of asthenospermia, and these markers may become the diagnostic and therapeutic targets of the disease.

[Identification of human testicular embryonal carcinoma proteins by two-dimensional gel electrophoresis and mass spectrometry].

OBJECTIVE: To separate and identify human testicular embryonal carcinoma proteomics using two-dimensional electrophoresis (2-DE) and mass spectrometry. METHODS: Immobilized pH gradient two-dimensional polyacrylamide gel electrophoresis was used to separate the total proteins of the samples. After silver staining, PDQuest 7.30 image analysis software was applied to analyze the 2-DE images. Three of the proteins highly expressed in human testicular embryonal carcinoma were identified by matrix-assisted laser adsorption/ionization-time of flight-tandem mass spectrometry (MALDI-TOF-MS/MS). RESULTS: 2-DE effectively screened the differentially expressed proteins in the carcinoma tissues. Three proteins highly expressed in the carcinoma were successfully identified. CONCLUSION: The proteins of human testicular embryonal carcinoma can be effectively separated and analyzed using 2-DE and mass spectrometry. Proteomic analysis offers a new means for further study of this carcinoma.

[Literature-mining and bioinformatic analysis of androgen-independent prostate cancer-specific genes].

OBJECTIVE: To compare the differences of the gene expressions in androgen-independent and androgen-dependent prostate cancer (ADPC), gain a deeper insight into the molecular mechanism of androgen-independent prostate cancer (AIPC), and find effective means for its clinical diagnosis and treatment. METHODS: Eats of genes highly-associated with prostate cancer were obtained by mining PubMed with the FACTA tool, and the specifically expressed genes in AIPC were analyzed with a set of bioinformatic tools including GATHER, PANTHER, STRING and ToppGene. RESULTS: A total of 128 genes specifically expressed in AIPC were identified, as compared with 23 that were specific to ADPC. Bioinformatic analysis showed the essential roles of AIPC-specific genes in such important biological processes as cell signal transduction, cell adhesion, apoptosis, oncogenesis, cell proliferation and cell differentiation. CONCLUSION: Such genes as MMPJ, EGFR, MMP2, ADM, MIF, IGFBP3, 112, MET, BAD, RHOA, SPP1, EP300, SMAD3, RAE1, PTK2, and TGFB2 may play important roles in transforming ADPC into AIPC.