Invasion Of DU145 Cells Research Articles

SU6668 modulates prostate cancer progression by downregulating MTDH/AKT signaling pathway.

Prostate cancer is the second leading cause of cancer deaths among men in Western counties and has increased in incidence also in China in recent years. Although diagnosis modalities for primary prostate cancer have markedly improved, there are still no effective therapies for metastatic prostate cancer. SU6668 is an inhibitor of the tyrosine kinase activity of three angiogenic receptors VEGFR2, PDGFRβ and FGFR1. There is strong experimental evidence that SU6668 can induce growth inhibition of various primary tumors. However, the function and molecular mechanism of SU6668 in prostate cancer has not been fully elucidated. In the present study, we found that SU6668 inhibited the proliferation and invasion of prostate cancer cells. Functional studies also demonstrated that SU6668 inhibited epithelial-mesenchymal transition in DU145 and LNCap cells. After treatment with SU6668, MTDH protein, which has been reported to be significantly overexpressed in many human tumor tissues, was downregulated in DU145 and LNCap cells. siRNA-mediated silencing of MTDH in prostate cancer cells decreased their proliferation and invasive capabilities, suggesting that SU6668 may inhibit cell proliferation and invasion of prostate cancer cells partly through downstream targeting of MTDH. Mechanistic investigations showed that AKT signaling pathway was inhibited after SU6668 treatment in prostate cancer cells. Moreover, a combination of SU6668 and PI3K-AKT pathway inhibitor LY29004 resulted in increased inhibition of cell proliferation and invasion in DU145 cells. Taken together, our findings revealed that SU6668 suppressed prostate cancer progression by downregulating MTDH/AKT signaling pathway and identified a promising therapeutic strategy for prostate cancer.

Inhibitory effect of oncolytic adenovirus ZD55-special AT rich sequence binding protein 1 on proliferation and invasion of andragon-independent prostate cancer DU145 cells

Objective To investigate the suppressive effect of oncolytic adenovirus ZD55-special AT rich sequence binding protein 1 (SATB1) expressing small hairpin RNA targeting SATB1 gene on growth and invasion of prostate cancer DU145 cells. Methods The oncolytic adenovirus ZD55-SATB1 was constructed and identifies. The inhibitory effect of ZD55-SATB1 on SATB1 expression was evaluated by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blotting analysis at 72 h after infection of DU145 cells with ZD55-SATB1 at a MOI of 10. The cytotoxicity of ZD55-SATB1 was detected by cell counting kit-8 (CCK-8) assay and crystal violet staining assay. Cell invasion was detected by Matrigel invasion assay. The expression of E1A protein in DU145 cell lines infected with viruses respectively was detected by Western blotting. Results The oncolytic adenoviruses ZD55-SATB1 were successfully constructed and identified. RT-PCR and Western blotting assay showed that ZD55-SATB1 could significantly decrease the expression of SATB1 in DU145 cells (P<0.05). The relative protein levels of E1A in DU145 cells treated with ZD55-SATB1 were significantly higher than those in human normal prostate epithelial cell line PZ-HPV-7 cells (P<0.01), which indicated cancer specificity of ZD55-SATB1 for prostate cance cells. Crystal violet staining assay showed that ZD55-SATB1 exhibited obvious cytotoxicity in DU145 cells. The cytopathic effect of ZD55-SATB1 was much stronger than that of ZD55-enhanced green fluorescent protein (EGFP) and Ad-SATB1. CCK-8 assay showed that cell viability of ZD55-SATB1 group [(31.60±3.31)%] was significantly lower than that in ZD55-EGFP group [(61.70±2.42)%, P=0.025] and Ad-SATB1 group [(82.40±3.54)%, P=0.017] for DU145 cells after 4 days. Transwell cell invasive experiment showed that the number of cells that invaded the membrane in ZD55-SATB1 group was (61.30±17.25) cells, significantly less than that in the Ad-SATB1 group [(170.10±25.44) cells, P=0.043], the ZD55-EGFP group [(295.40±23.36) cells, P=0.013] and the PBS group [(490.50±33.36) cells, P=0.009]. Conclusion Oncolytic adenovirus ZD55-SATB1 could effectively knockdown the SATB1 expression in DU145 cells, and dramatically inhibit proliferation and invasion of DU145 cells. Key words: Oncolytic adenovirus; Special AT rich sequence binding protein 1; Proliferation; Invasion

Upregulation of N - myc downstram regulated gene 1 expression by silencing DNA methyltransferases influences the biological behaviors of prostate cancer DU145 cells

Objective To investigate the influence of the upregulation of N-myc downstram regulated gene 1 (NDRG1) expression by silencing DNA methyltransferases (DNMTs) using small interfering RNAs (siRNAs) on the biological behaviors of prostate cancer DU145 cells. Methods SiRNAs were used to inhibit the expression of DNMT1, DNMT3b of DU145 cells; Western blotting and quantitative real-time reverse transcriptase-polymerase chain reaction (RT-qPCR) were used to detect the expression levels of DNMT1, DNMT3b and NDRG1 of DU145 cells; cell counting kit-8 (CCK-8) assay, muse cell analyzer, wound healing assay and Transwell migration assay were used to detect the proliferation, apoptosis, migration and invasion of DU145 cells; then we inhibited the expression of NDRG1 of DU145 cells which had the most significant changes of cell biological behaviors; and we detected the expression level of NDRG1 and the changes of cell biological behaviors. Results Western blotting and RT-qPCR showed that the mRNA (1.59±0.03, 1.23±0.01, 1.36±0.11 vs.0.90±0.15, 1.00±0.03) and protein (1.07±0.16, 0.49±0.01, 0.92±0.09 vs.0.23±0.04, 0.30±0.07) expression levels of NDRG1 of DNMT1 inhibition group, DNMT3b inhibition group and double inhibition group were significantly higher than these in control groups (P<0.05). CCK-8 assay showed that, at 24, 48 and 72 h, the optical density (A) values of DNMT1 inhibition group (0.189±0.030, 0.266±0.048, 0.419±0.058), DNMT3b inhibition group (0.221±0.023, 0.318±0.072, 0.498±0.064) and double inhibition group (0.199±0.021, 0.284±0.062, 0.488±0.091) were significantly lower than control groups (P<0.05). Apoptosis results showed that the apoptosis rates of DNMT1 inhibition group (22.40±1.36)%, DNMT3b inhibition group (14.20±0.80)% and double inhibition group (17.60±0.74)% were significantly higher than control groups (P<0.05). Wound healing assay showed that the healing rates of DNMT1 inhibition group (53.01±3.22)%, DNMT3b inhibition group (70.97±2.12)% and double inhibition group (63.93±4.42)% were significantly lower than control groups (P<0.05). Transwell migration assay showed that the amount of invasion cells in DNMT1 inhibition group [(16±3) cells], in DNMT3b inhibition group [(32±5) cells] and in double inhibition group [(20±4) cells] were significantly less than control groups (P<0.05). Among these groups, DNMT1 inhibition group had the most significant effect (P<0.05), but the co-transfection had no significant synergistic effect. Compared with control groups, NDRG1-siRNA could significantly reverse the changes of cell biological behaviors of DU145 cells (P<0.05). Conclusion Inhibition of DNMT1, DNMT3b of DU145 cells could partially restore NDRG1 expression, which could inhibit the proliferation, apoptosis, migration and invasion of cancer cells. Moreover, DNMT1 inhibition group had the most significant effect, but the co-transfection had no significant synergistic effect, suggesting that DNMT1 could become an effective target for demethylation treatment of prostate cancer. Key words: N-myc downstram regulated gene 1; DNA methylation; DNA methyltransferase; RNA interference; Prostate cancer

MicroRNA-372 Suppresses Migration and Invasion by Targeting p65 in Human Prostate Cancer Cells.

Prostate cancer (PCa) is one of the most prevalent malignant tumors. microRNAs (miRNAs) play an important role in cancer initiation, progression, and metastasis, and their roles in PCa are becoming more apparent. In this study, we found that microRNA-372 (miR-372) is downregulated in human PCa and inhibits the proliferation activity, migration, and invasion of DU145 cells. Subsequently, p65 is confirmed as a target of miR-372, and knockdown of p65 expression similarly resulted in decreased proliferation activity, migration, and invasion. CDK8, MMP-9, and prostate-specific antigen were involved in both these processes. Taken together, our results show evidence that miR-372 may function as a tumor suppressor gene by regulating p65 in PCa and may provide a strategy for blocking PCa metastasis.

Metformin Elicits Antitumor Effects and Downregulates the Histone Methyltransferase Multiple Myeloma SET Domain (MMSET) in Prostate Cancer Cells.

This study explored the biological effects of metformin on prostate cancer (PCa) cells and determined molecular pathways and epigenetic regulators implicated in its mechanism of action. We performed mRNA expression profiling in 22Rv1 cells following 2.5 mM and 5 mM metformin treatment. Genes significantly modified by metformin treatment were ranked based on altered expression, involvement with cancer-related processes, and reported dysregulation in PCa. The effects of the top ranked gene, MMSET, on the proliferative and invasive capabilities of PCa cells were investigated via siRNA knockdown alone and also combined with metformin treatment. Metformin treatment decreased cell growth of PCa cell line 22Rv1 and stalled cells at the G1/S checkpoint in a time- and dose-dependent manner, resulting in increased cells in G1 (P < 0.05) and decreased cells in S (P < 0.05) phase. Metformin activated the AMPK/mTOR signaling pathway as shown by increased p-AMPK and decreased p-p70S6K. mRNA expression profiling following metformin treatment identified significant changes in 136 chromatin-modifying genes. The top ranked gene, multiple myeloma SET domain (MMSET) showed increased expression in PCa cell lines (22Rv1 and DU145) when compared to the benign prostate epithelium-derived cell-line RWPE-1, and its expression was decreased upon metformin treatment. siRNA-mediated knockdown of MMSET showed decreased cellular migration and invasion in DU-145 cells. MMSET knockdown in combination with metformin treatment resulted in further reduction in the capacity of PCa cells to migrate and invade. These data suggest MMSET may play a role in the inhibitory effect of metformin on PCa and could serve as a potential novel therapeutic target for PCa. Prostate 76:1507-1518, 2016. © 2016 Wiley Periodicals, Inc.

Simvastatin blocks TGF-β1-induced epithelial-mesenchymal transition in human prostate cancer cells.

In recent years, the use of statins has been reported to be associated with a reduced risk of prostate cancer (PCa), particularly metastatic PCa. The mechanisms underlying these epidemiological observations are poorly understood. Epithelial-mesenchymal transition (EMT) is a critical initial step and a hallmark for cancer metastasis. In the present study, the relationship between simvastatin and EMT in PCa and the mechanism involved was investigated. It was demonstrated that simvastatin inhibited the EMT as assessed by reduced expression of N-cadherin and vimentin, and increased E-cadherin in TGF-β1 treated DU145 PCa cells. Furthermore, simvastatin inhibited TGF-β1-induced migration and invasion of DU145 cells. The TGF-β1/Smad pathway and non-Smad pathway were investigated in simvastatin-treated DU145 cells. Simvastatin had no effect on TGF-β1-induced phosphorylation of Smad2 and Smad3. In the non-Smad pathway, simvastatin reduced TGF-β1-induced p38 MAPK phosphorylation, but had no effect on TGF-β1-induced Erk1/2 phosphorylation. Simvastatin attenuated TGF-β1-induced EMT, cell migration and invasion in DU145 cells. These effects may have been mediated by the inhibition of p38 MAPK phosphorylation, not through the canonical Smad pathway. Therefore simvastatin may be a promising therapeutic agent for treating PCa.

Anti-tumor activity of the TRPM8 inhibitor BCTC in prostate cancer DU145 cells

The present study investigated the anti-tumor activity of N-(4-tertiarybutylphenyl)-4-(3-chloropyridin-2-yl)tetrahydropyrazine-1(2H)-carbox-amide (BCTC), a potent and specific inhibitor of transient receptor potential cation channel subfamily M member 8 (TRPM8) in prostate cancer (PCa) DU145 cells. TRPM8 expression in DU145 and normal prostate PNT1A cells was detected by reverse transcription polymerase chain reaction and western blot analysis. The effect of BCTC on DU145 cells was analyzed by flow cytometry analysis, and MTT, scratch motility and Transwell invasion assays. The molecular mechanism through which BCTC acts was investigated by western blot analysis. TRPM8 expression was increased in DU145 cells compared with PNT1A cells at the mRNA and protein levels. The present study provided evidence that inhibition of TRPM8 by BCTC reduced the viability of DU145 cells, but not PNT1A cells. In addition, BCTC inhibited cell cycle progression, migration and invasion in DU145 cells. Cell cycle-associated proteins, including phosphorylated protein kinase B, cyclin D1, cyclin dependent kinase (CDK) 2 and CDK6 were downregulated by BCTC, while phosphorylated glycogen synthase kinase 3β was upregulated. However, investigations in the present study revealed that BCTC failed to trigger apoptosis in DU145 cells. In addition, in BCTC-treated DU145 cells, phosphorylated extracellular signal-regulated kinase 1/2 was downregulated substantially while phosphorylated p38 (p-p38) and phosphorylated c-Jun N-terminal kinases (p-JNK) were upregulated. The anti-proliferative activity of BCTC on DU145 cells was attenuated by p38 and JNK-specific inhibitors, suggesting that MAPK pathways are involved. Overall, the TRPM8 specific antagonist BCTC demonstrated excellent anti-tumor activity in PCa DU145 cells, and therefore has the potential to become a targeted therapeutic strategy against PCa.

Camptothecin suppresses expression of matrix metalloproteinase-9 and vascular endothelial growth factor in DU145 cells through PI3K/Akt-mediated inhibition of NF-κB activity and Nrf2-dependent induction of HO-1 expression

Though camptothecin (CPT) possesses potent anti-inflammatory, immunomodulatory, anticancerous, and antiproliferative effects, little is known about the mechanism by which CPT regulates the expression of matrix metalloproteinase-9 (MMP-9) and vascular endothelial growth factor (VEGF). Therefore, the current study aimed to investigate the effects of CPT on the expression of MMP-9 and VEGF, which are important factors for the invasion of tumors. In vitro application of CPT resulted in a slight inhibition of cell proliferation and a significant reduction in the matrigel invasion of DU145 cells. Treatment with CPT also downregulated phorbol-12-myristate-13-acetate (PMA)- and tumor necrosis factor-α (TNF-α)-induced MMP-9 and VEGF expression by inhibiting nuclear factor-κB (NF-κB) activity. Downregulation of phosphoinositide 3-kinase (PI3K)/Akt phosphorylation in response to CPT was revealed as an upstream pathway regulating the expression of MMP-9 and VEGF accompanying the inhibition of NF-κB activity. We further confirmed that CPT inhibits PMA-induced MMP-9 and VEGF expression by upregulating nuclear factor-erythroid related factor-2 (Nrf2)-mediated heme oxygenase-1 (HO-1) induction. Taken together, these data indicate that CPT inhibits the invasion of cancer cells accompanied by suppression of MMP-9 and VEGF production by suppressing the PI3K/Akt-mediated NF-κB pathway and enhancing the Nrf2-dependent HO-1 pathway, suggesting that CPT may be a good candidate to inhibit MMP-9 and VEGF expression.

P0065 S100A16 promotes cell migration and invasion in prostate cancer in vitro via AKT and ERK cell signalling pathways

<h3>Background</h3> Expression of S100A16, a member of the S100 calcium binding protein family, is elevated in many types of tumours and appears to be associated with tumour cell proliferation. <h3>Methods</h3> By use of human prostate cancer cells as the model system, we evaluated the functional roles and underlying pathways of S100A16 in the proliferation and migration of prostate cancer cells. <h3>Findings</h3> S100A16 was significantly overexpressed in both human prostate cancer tissues and cell lines compared with normal controls. Exogenously introduced overexpression of S100A16 significantly promoted invasion, migration, and proliferation in human prostate cancer cells in vitro; whereas silencing the expression of S100A16 showed the opposite effects. Furthermore, overexpression of S100A16 activated AKT and ERK and reduced levels of the tumour suppressors p21 and p27. Pharmacological inhibition of PI3-kinase and MAP kinase via LY294002 and PD98059, respectively, attenuated the clone formation and invasion of DU-145 cells induced by S100A16 overexpression. <h3>Interpretation</h3> Our findings suggest that S100A16 can promote human prostate cancer progression via AKT and ERK signalling pathways as well as cell cycle regulators p21 and p27. S100A16 may serve as a novel therapeutic target or diagnostic marker of human prostate cancer progression.

SATB1 is overexpressed in metastatic prostate cancer and promotes prostate cancer cell growth and invasion

BackgroundSpecial AT-rich sequence binding protein 1 (SATB1) is a nuclear factor that functions as the global chromatin organizer to regulate chromatin structure and gene expression gene expression. SATB1 has been shown to be abnormally expressed in various types of cancer. However, the expression and role of SATB1 in prostate cancer remain unclear.Methods120 cases of prostatic carcinoma and 60 cases of benign prostate hyperplasia were analyzed for SATB1 expression by immunohistochemistry. LNCaP, DU-145, and PC3 prostate cancer cells were examined for SATB1 expression by Western blot analysis. Cell proliferation and invasion was evaluated by CCK8 and transwell invasion assay, respectively.ResultsSATB1 staining was stronger in prostatic carcinomas with metastasis than in those without metastasis, but was absent in benign prostate hyperplasia. Furthermore, SATB1 expression was positively correlated with bone metastasis and the Gleason score. SATB1 overexpression promoted the proliferation and invasion of LNCaP cells while SATB1 knockdown inhibited the proliferation and invasion of DU-145 cells.ConclusionsThese findings provide novel insight into oncogenic role of SATB1 in prostate cancer, suggesting that SATB1 is a promising biomarker and therapeutic target for prostate cancer.

Identification of cetrimonium bromide and irinotecan as compounds with synthetic lethality against NDRG1 deficient prostate cancer cells

The N-myc downstream regulated gene 1 (NDRG1) has been identified as a metastasis-suppressor gene in prostate cancer (PCa). Compounds targeting PCa cells deficient in NDRG1 could potentially decrease invasion/metastasis of PCa. A cell based screening strategy was employed to identify small molecules that selectively target NDRG1 deficient PCa cells. DU-145 PCa cells rendered deficient in NDRG1 expression by a lentiviral shRNA-mediated knockdown strategy were used in the primary screen. Compounds filtered from the primary screen were further validated through proliferation and clonogenic survival assays in parental and NDRG1 knockdown PCa cells. Screening of 3360 compounds revealed irinotecan and cetrimonium bromide (CTAB) as compounds that exhibited synthetic lethality against NDRG1 deficient PCa cells. A three-dimensional (3-D) invasion assay was utilized to test the ability of CTAB to inhibit invasion of DU-145 cells. CTAB was found to remarkably decrease invasion of DU-145 cells in collagen matrix. Our results suggest that CTAB and irinotecan could be further explored for their potential clinical benefit in patients with NDRG1 deficient PCa.

The Role of the Protein C and Urokinase Systems in Prostate Cancer

The objective of this study was to determine how the protein C and urokinase systems regulate prostate tumor cell migration. Prostate cancer (CaP) is the most frequently diagnosed cancer among males, with an estimated 238,590 new cases arising in the United States during 2013. Urokinase plasminogen activator (uPA) and its inhibitor, plasminogen activator inhibitor‐1 (PAI‐1), are regulators of blood coagulation and normal cell migration. Abnormal secretion of these two proteins by neoplastic cells in the tumor microenvironment has been linked to an increased likelihood of (CaP) invasion. Using a modified Boyden chamber, we tested the ability of activated protein C (APC), uPA, PAI‐1 and vitronectin (VN), an ECM protein, to mediate uPA and PAI‐1 dependent migration of the DU‐145 CaP cell line through Matrigel. As has been shown for ovarian cancer, in the presence of uPA and PAI‐1, APC competed with uPA for binding to PAI‐1, enabling uPA to increase the invasiveness of DU‐145 cells. We also determined that VN acts as a cofactor that accelerates APC‐PAI‐1 interaction, potentially allowing once inhibited uPA to degrade ECM and give neoplastic cells room to invade through tissue. We conclude that although protein C and APC alone do not affect CaP cell invasion, in the presence of uPA and PAI‐1, APC binds to PAI‐1, freeing uPA to facilitate CaP tumor cell invasion.

Crude extract of Euphorbia formosana inhibits the migration and invasion of DU145 human prostate cancer cells: The role of matrix metalloproteinase-2/9 inhibition via the MAPK signaling pathway

Prostate cancer is a common worldwide health problem in males with a poor prognosis due in part to tumor invasion and migration. The crude extract of Euphorbia formosana (CEEF) has been used for the treatment of numerous diseases, however, its effects on the migration and invasion of prostate cancer cells have yet to be examined. In the present study, we investigated the effects of CEEF on the migration and invasion of DU145 human prostate cancer cells invitro. The wound healing assay and the Matrigel-uncoated migration assay were used to examine the migration of cancer cells. Western blotting was used to examine the levels of proteins associated with migration and invasion, and gelatin zymography was used to examine the secretion levels of matrix metalloproteinases-2 and -9 (MMP‑2/9) from DU145 cells following exposure to CEEF. The results indicated that CEEF suppressed the migration and invasion of DU145 prostate cancer cells and that these effects are exerted in a concentration- and time-dependent manner. CEEF inhibited the ERK1/2, p38, JNK, SOS1, PKC, PI3K and MMP-2/9 protein expression in DU145 cells. The results demonstrated that CEEF suppressed the migration and invasion of DU145 cells through inhibition of the mitogen-activated protein kinase (MAPK) signaling pathway resulting in the inhibition of MMP-2/9 in DU145 human prostate cancer cells.

Piceatannol inhibits MMP-9-dependent invasion of tumor necrosis factor-α-stimulated DU145 cells by suppressing the Akt-mediated nuclear factor-κB pathway

Piceatannol has potent anti-inflammatory, immunomodulatory, anticancer and antiproliferative effects. However, little is known about the mechanism by which piceatannol inhibits invasion and metastasis. The aim of the current study was to investigate the effects of piceatannol on the expression of matrix metalloproteinase-9 (MMP-9) in DU145 human prostate cancer cells. The results revealed that MMP-9 activity was significantly increased in response to tumor necrosis factor-α (TNF-α). However, treatment with piceatannol reversed TNF-α- and MMP-9-induced gelatin zymography and its gene expression. In addition, a Matrigel invasion assay determined that piceatannol reduces the TNF-α-induced invasion of DU145 cells. Nuclear factor-κ B (NF-κB) is a significant transcription factor that regulates numerous genes involved in tumor cell invasion and metastasis. Therefore, whether piceatannol acts on NF-κB to regulate MMP-9 gene expression was analyzed. The results revealed that piceatannol attenuates MMP-9 gene expression via the suppression of NF-κB activity. Using a specific NF-κB inhibitor, pyrrolidine dithiocarbamate, it was confirmed that TNF-α-induced MMP-9 gene expression is primarily regulated by NF-κB activation. Piceatannol inhibited NF-κB activity by suppressing nuclear translocation of the NF-κB p65 and p50 subunits. Furthermore, TNF-α-induced Akt phosphorylation was significantly downregulated in the presence of piceatannol. The Akt inhibitor LY294002 caused a significant decrease in TNF-α-induced NF-κB activity and MMP-9 gene expression. Overall, these data suggest that piceatannol inhibits TNF-α-induced invasion by suppression of MMP-9 activation via the Akt-mediated NF-κB pathway in DU145 prostate cancer cells.

Hypoxia increases CX3CR1 expression via HIF-1 and NF-κB in androgen-independent prostate cancer cells

The unique CX3C chemokine CX3CL1 and its cognate receptor CX3CR1 have been implicated in organ-specific metastasis of various types of tumors. Hypoxia, a common phenomenon in solid tumors, is associated with a malignant cancer phenotype. Previous studies have proved that hypoxia facilitates cancer cell metastasis through upregulation of specific chemokine receptors. We hypothesized that hypoxia could upregulate CX3CR1 expression and lead to an increased chemotactic response to CX3CL1 in prostate cancer cells. In the present study, we found that CX3CR1 expression was significantly increased in androgen-independent prostate cancer cells, including DU145, PC-3 and PC-3M, following exposure to hypoxia. This upregulation of CX3CR1 corresponded to a significant increase in migration and invasion of prostate cancer cells under hypoxic conditions, which was attenuated after knocking down CX3CR1 expression. In addition, we examined the possible role of HIF-1 and NF-κB in the process of hypoxia-induced CX3CR1 expression and hypoxia-mediated metastasis. Attenuation of HIF-1 and NF-κB transcriptional activity by siRNAs or pharmacological inhibitors, abrogated hypoxia-induced upregulation of CX3CR1, and also prevented the migration and invasion of DU145 cells under a hypoxic environment. In summary, our study demonstrated that HIF-1 and NF-κB are essential for hypoxia-regulated CX3CR1 expression, which is associated with increased migratory and invasive potential of prostate cancer cells. CX3CR1 signaling is a potential therapeutic target in the adjuvant treatment of prostate cancer.

Effects of oldhamianoside II on the invasive ability of prostatic carcinoma cells

Objective To observe the effects of oldhamianoside Ⅱ on the invasive 'ability of prostatic carcinoma DU-145 cells,and to investigate the potential mechanism.Methods After treatment of DU-145 cells with oldhamianoside Ⅱ,MTT assay was used to test the effect of oldhamianoside Ⅱ on the cell proliferation.The invasive ability was assessed with a transwell cell culture chamber.Immunocytochemistry stain was used to investigate the expression of matrix metalloproteinase(MMP-2)and CXC-chemokine receptor 4 (CXCR4).Results The proliferation of DU-145 cells was inhibited after treatment with oldhamianoside Ⅱ.In vitro invasion assay indicated that the cells moved through the membrane were(180.3 ± 14.6)/field in the control group,while decreased to(100.4 ±1.5),(80.2±2.7)and(60.1 ±5.3)/field in 0.25,0.5 and 1μg/mL oldhamianoside Ⅱ treated DU-145 cells respectively.Comparing to control group,the cells of oldhamianoside Ⅱ groups moved through the membrane were decreased remarkably(P ＜0.05).Immunocytochemistry showed that the expression of MMP-2 and CXCR4 were decreased of oldhamianoside Ⅱ groups.Conclusions Oldhamianoside Ⅱ can inhibit the invasiveness of DU-145 cells,which is related to the down regulation the expression of MMP-2 in DU-t45 cells.Oldhamianoside Ⅱ can inhibit the prostate cancer cells metastasis to the bone,which is related to the down regulation the expression of CXCR4 in DU-145 cells. Key words: Prostatic neoplasms; Matrix metalloproteinases; Receptors; CXCR4; Oldhamianoside Ⅱ

782 SANGUINARINE INHIBITS STAT3 ACTIVATION AND TUMOR GROWTH IN PROSTATE CANCER CELLS

You have accessJournal of UrologyProstate Cancer: Basic Research IV1 Apr 2012782 SANGUINARINE INHIBITS STAT3 ACTIVATION AND TUMOR GROWTH IN PROSTATE CANCER CELLS Meng Sun, Chengfei Liu, Wei Lou, Nagalakshmi Nadiminty, Joy Yang, Christopher Evans, and Allen Gao Meng SunMeng Sun Sacramento, CA More articles by this author , Chengfei LiuChengfei Liu Sacramento, CA More articles by this author , Wei LouWei Lou Sacramento, CA More articles by this author , Nagalakshmi NadimintyNagalakshmi Nadiminty Sacramento, CA More articles by this author , Joy YangJoy Yang Sacramento, CA More articles by this author , Christopher EvansChristopher Evans Sacramento, CA More articles by this author , and Allen GaoAllen Gao Sacramento, CA More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2012.02.870AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Signal Transducer and Activator of Transcription 3 (STAT3) is an oncogenic transcription factor implicated in prostate carcinogenesis and progression. Stat3 activation is associated with prostate cancer development and progression. Constitutively activated Stat3 activates androgen receptor signaling and promotes castration-resistant prostate cancer growth. Targeting Stat3 activation is an effective strategy in prostate cancer therapy. METHODS DU145, C4-2B and LNCaP cells were treated with sanguinarine for 4 hours and then stimulated with Interleukin-6 (IL-6) or vehicle. The phosphorylation status of STAT3 and related proteins were measured with western blots. Activation of transcription by STAT3 via the STAT3 responsive element was measured with luciferase reporter assay. The effect of sanguinarine on DU145 and LN/S17 cells anchorage-independent growth were examined with soft agar assay. The effect of sanguinarine on cells migration and invasion of DU145 cells were measured with scratch assay and invasion assay respectively. RESULTS Exposure to micromolar concentrations of sanguinarine resulted in suppression of constitutive as well as IL6-induced phosphorylation of STAT3 at both Tyr705 and Ser727 in DU145 cells. The inhibition of STAT3 phosphorylation correlated with the inhibition of Janus-activated Kinase 2 (JAK2) and src phosphorylation. Sanguinarine inhibited the constitutive and IL6-induced STAT3 responsive element luciferase activities. Several STAT3 regulated genes such as c-myc and survivin were downregulated by sanguinarine. We found that sanguinarine inhibited the migration and invasion of DU145 cells that express constitutively activated STAT3. Furthermore, sanguinarine inhibited the anchorage-independent growth of DU145 and LN/S17 cells and suppressed the growth of DU145 xenografts in nude mice. CONCLUSIONS These data suggest that sanguinarine targets multiple upstream kinases that mediate STAT3 activity. Sanguinarine is a potent STAT3 inhibitor that could be developed as a therapeutic agent for prostate cancer. © 2012 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 187Issue 4SApril 2012Page: e320 Advertisement Copyright & Permissions© 2012 by American Urological Association Education and Research, Inc.MetricsAuthor Information Meng Sun Sacramento, CA More articles by this author Chengfei Liu Sacramento, CA More articles by this author Wei Lou Sacramento, CA More articles by this author Nagalakshmi Nadiminty Sacramento, CA More articles by this author Joy Yang Sacramento, CA More articles by this author Christopher Evans Sacramento, CA More articles by this author Allen Gao Sacramento, CA More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

IJU this issue

This issue consists of two Perspectives in Urology, three Review Articles, five Original Articles, one Case Report, one Short Communication and two other reports. The content of this issue focuses on various interesting hot topics associated with clinical, as well as basic, aspects; therefore, I believe that this issue might help provide useful information for a wide variety of readers of IJU. This issue starts with Perspectives in Urology, a newly introduced category in IJU, which briefly presents a scientific view of several fields of urology by specialists. Two articles published in this issue describe the application of gasless single-port surgery to partial nephrectomy by Kihara (Tokyo, Japan) and controversial issues in the decontamination process for urological endoscopes by Yanaihara et al. (Saitama, Japan). This issue then includes three interesting Review Articles. Takenaka et al. (Tottori, Japan) summarized the recent progress in anatomical findings in relation to each basic step during robotic-assisted laparoscopic radical prostatectomy. They emphasized the important message that surgical anatomy should always be developing and changing with advances in surgical approaches. Matsumoto et al. (Asahikawa, Japan) described the pathophysiological relationship between metabolic syndrome and lower urinary tract symptoms (LUTS) with a special emphasis on bladder blood flow. They also proposed a novel therapeutic strategy for patients with LUTS from the viewpoint of bladder blood flow. Azuma et al. (Takatsuki, Japan) reviewed their experience of the development of novel bladder preservation approach for patients with locally advanced bladder cancer. They carried out combined therapy using balloon-occluded arterial infusion anticancer agents and hemodialysis with concurrent radiation, and presented promising oncological outcomes. Consequently, five excellent Original Articles are provided. Jensen et al. (Aarhus, Denmark) compared the prognostic significance between extended and limited lymph node dissection in patients with bladder cancer who underwent radical cystectomy. Although extended nodal dissection provided more accurate staging than limited dissection, recurrence patterns appeared to be similar between these two groups. Shigemura et al. (Kobe, Japan) compared Wallace direct ureteroileal anastomosis with the Le Duc anti-reflux procedure in modified Studer orthotopic neobladder reconstruction after radical cystectomy. The authors concluded that direct ureteroileal anastomosis using the Wallace method is effective for minimizing the stenosis of ureteroileal anastomosis. Bayraktar et al. (Istanbul, Turkey) assessed the effect of constipation on serum prostate-specific antigen (PSA) levels in men. In this study, serum PSA values in the constipated patients before treatment were shown to be significantly higher than those in the control group; therefore, the authors recommended the re-evaluation of PSA value in the constipated patients before carrying out the prostate biopsy. Ip et al. (Taichung, Taiwan) reported the apoptotic effects of bee venom on human bladder cancer cells and the signaling pathways involved in this event. This article clearly showed the induction of both caspase-dependent and caspase-independent apoptotic cell death in human bladder cancer cells by bee venom through the intracellular Ca2+-modulated intrinsic death pathway. Nakane et al. (Gifu, Japan) analyzed the role of cortactin and silent mating type information regulation 2 homolog 1 (SIRT1) in migration and invasion of human prostate cancer DU145 cells. Because knockdown of cortaction or SIRT1 expression resulted in the inhibition of both migration and invasion of DU145 cells, the authors suggest that this strategy could be a promising treatment for prostate cancer by regulating its metastatic spread. This issue also included an interesting Case Report and Short Communication. Fujisaki et al. (Shimotsuke-city, Japan) reported life-threatening anaphylaxis to leuprorelin acetate depot. Sugihara et al. (Iwata, Japan) reported the low incidence of admissions related to interstitial cystitis in Japan, which could possibly be explained by several factors, including racial difference, examination methods, lack of outpatient data and poor health-care coverage of this disease. None declared.

Inhibition of cortactin and SIRT1 expression attenuates migration and invasion of prostate cancer DU145 cells

Cortactin is overexpressed in various types of cancer and enhances cell motility. It has been recently reported that silent mating type information regulation 2 homolog 1 interacts with cortactin and promotes cell migration. Here, we examined the role of cortactin and silent mating type information regulation 2 homolog 1 in migration and invasion of prostate cancer cells. The cortactin expression levels in DU145, LNCaP and PC3 prostate cancer cells, and in PrEC normal human prostate epithelial cells were evaluated by western blot analysis. In DU145 cells, the expression of cortactin or silent mating type information regulation 2 homolog 1 was inhibited by small interfering RNA, and the effects of their knockdown on migration and invasion were examined by cell migration and invasion assays. To determine the localization of cortactin and silent mating type information regulation 2 homolog 1, western blot and immunofluorescence microscopic analyses were carried out. The functional interaction between silent mating type information regulation 2 homolog 1 and cortactin was also studied by in vivo acetylation assay. The protein expression of cortactin was significantly higher in DU145 cells than in other cell lines. Knockdown of cortactin or silent mating type information regulation 2 homolog 1 expression inhibited both migration and invasion of DU145 cells. Similarly to cortactin, silent mating type information regulation 2 homolog 1 was found to be predominantly expressed in the cytoplasm. Finally, the knockdown of silent mating type information regulation 2 homolog 1 expression increased the acetylation level of cortactin. Our findings suggest that inhibition of cortactin or silent mating type information regulation 2 homolog 1 expression attenuates migration and invasion of DU145 cells and this could represent a promising strategy to regulate metastasis of prostate cancer.

Glionitrin B, a Cancer Invasion Inhibitory Diketopiperazine Produced by Microbial Coculture

A new diketopiperazine, glionitrin B (1), was produced using a microbial coculture of the fungus Aspergillus fumigatus KMC-901 and the bacterium Sphingomonas sp. KMK-001 that were isolated from acidic coal mine drainage. The structure of 1 was determined to be (3S,10aS)-dithiomethylglionitrin A. This structure was determined by the analyses of extensive NMR data and the circular dichroism spectra of the natural product and a semisynthetic compound derived from glionitrin A. In contrast to glionitrin A (2), glionitrin B (1) is not cytotoxic against the human prostate cancer cell line DU145. However, compound 1 caused suppression of DU145 cell invasion, producing 46% inhibition at 60 μM.