Urological Cancer Cell Research Articles

Enhancing Urological Cancer Treatment: Leveraging Vasodilator Synergistic Potential with 5-FU for Improved Therapeutic Outcomes.

Backgroud: This study investigates the potential of vasodilator drugs as additive therapy in the treatment of urological cancers, particularly in combination with the antineoplastic agent 5-fluorouracil (5-FU). Methods: The study evaluated the cytotoxic effects of sildenafil, tezosentan and levosimendan alone and in combination with 5-FU on urological cancer cell lines. The assessment included MTT assays, colony formation assays and wound healing assays to determine cell viability, proliferative capacity, and migratory behavior, respectively. Results: Sildenafil and tezosentan showed limited cytotoxic effects, while levosimendan demonstrated moderate anticancer activity. The combination of levosimendan and 5-FU exhibited an additive interaction, enhancing cytotoxicity against cancer cells while sparing normal cells. Levosimendan also inhibited cell migration and proliferation, potentially through mechanisms involving the modulation of cAMP levels and nitric oxide production. Conclusions: The findings suggest that levosimendan can be used in conjunction with 5-FU to reduce the required dose of 5-FU, thereby minimizing side effects without compromising therapeutic efficacy. This study offers a new perspective for enhancing therapeutic outcomes in patients with urological cancers.

Integrins are enriched on aberrantly fucosylated tumour‐derived urinary extracellular vesicles

AbstractUrinary extracellular vesicles (uEVs) are enriched with glycosylated proteins which have been extensively studied as putative biomarkers of urological cancers. Here, we characterized the glycosylation and integrin profile of EVs derived from urological cancer cell lines. We used fluorescent europium‐doped nanoparticles coated with lectins and antibodies to identify a biomarker combination consisting of integrin subunit alpha 3 (ITGA3) and fucose. In addition, we used the same cancer cell line‐derived EVs as analytical standards to assess the sensitivity of the ITGA3‐UEA assay. The clinical performance of the ITGA3‐UEA assay was analysed using urine samples of various urological pathologies including diagnostically challenging benign prostatic hyperplasia (BPH), prostate cancer (PCa) and bladder cancer (BlCa). The assay can significantly discriminate BlCa from all other patient groups: PCa (9.2‐fold;p = 0.00038), BPH (5.5‐fold;p = 0.004) and healthy individuals (and 23‐fold;p = 0.0001). Our results demonstrate that aberrantly fucosylated uEVs and integrin ITGA3 can be detected with fucose‐specific lectin UEA in a simple bioaffinity assay for the detection of BlCa directly from unprocessed urine.

BladMetrix: a novel urine DNA methylation test with high accuracy for detection of bladder cancer in hematuria patients

BackgroundCystoscopy is the gold standard for bladder cancer detection, but is costly, invasive and has imperfect diagnostic accuracy. We aimed to identify novel and accurate DNA methylation biomarkers for non-invasive detection of bladder cancer in urine, with the potential to reduce the number of cystoscopies among hematuria patients.ResultsBiomarker candidates (n = 32) were identified from methylome sequencing of urological cancer cell lines (n = 16) and subjected to targeted methylation analysis in tissue samples (n = 60). The most promising biomarkers (n = 8) were combined into a panel named BladMetrix. The performance of BladMetrix in urine was assessed in a discovery series (n = 112), consisting of bladder cancer patients, patients with other urological cancers and healthy individuals, resulting in 95.7% sensitivity and 94.7% specificity. BladMetrix was furthermore evaluated in an independent prospective and blinded series of urine from patients with gross hematuria (n = 273), achieving 92.1% sensitivity, 93.3% specificity and a negative predictive value of 98.1%, with the potential to reduce the number of cystoscopies by 56.4%.ConclusionsWe here present BladMetrix, a novel DNA methylation urine test for non-invasive detection of bladder cancer, with high accuracy across tumor grades and stages, and the ability to spare a significant number of cystoscopies among patients with gross hematuria.

Organ-Specific Uptake of Extracellular Vesicles Secreted by Urological Cancer Cells.

Simple SummaryExtracellular vesicles (EVs) play an important role in the communication of cancer cells with their local microenvironment and distant organ systems, in order to promote a supportive tumor microenvironment, as well as to prepare premetastatic niches. In this study, we aimed to analyze if the EVs secreted by urological cancer cells are taken up by specific organ systems, depending on their origin. After the intravenous injection of fluorescence-labeled EVs from benign and malignant prostate, kidney, and bladder cells in immunodeficient mice, their organs were harvested and analyzed for the presence of fluorescent EVs. We could show that (i) EVs are taken up not entirely organ-specifically but in different amounts, depending on their origin; (ii) EVs from malignant cells are taken up more efficiently than EVs from benign cells; and (iii) EVs are taken up very fast. These observations hint to an organotropism in EV uptake, which needs to be further investigated.Extracellular vesicles (EVs) secreted by cancer cells have been shown to take a pivotal part in the process of local and systemic tumor progression by promoting the formation of a supportive local tumor microenvironment and preparing premetastatic niches in distant organ systems. In this study, we analyzed the organ-specific uptake of EVs secreted by urological cancer cells using an innovative in-vivo approach. EVs from benign and malignant prostate, kidney, and bladder cells were isolated using ultracentrifugation, fluorescence-labeled and injected intravenously in immunodeficient mice. After 12 or 24 h, the animals were sacrificed, their organs were harvested and analyzed for the presence of EVs by high-resolution fluorescence microscopy. Across all entities, EVs were taken up fast (12 h > 24 h), and EVs from malignant cells were taken up more efficiently than EVs from benign cells. Though not entirely organ-specific, EVs were incorporated in different amounts, depending on the entity (prostate: lung > liver > brain; kidney: brain > lung > liver; bladder: lung > liver > brain). EV uptake in other organs than lung, liver, brain, and spleen was not observed. Our results suggest a role of EVs in the formation of premetastatic niches and an organotropism in EV uptake, which have to be examined in more detail in further studies.

Urinary exosomes-based Engineered Nanovectors for Homologously Targeted Chemo-Chemodynamic Prostate Cancer Therapy via abrogating EGFR/AKT/NF-kB/IkB signaling

Multi-functional nanovectors based on exosomes from cancer cell culture supernatants in vitro has been successfully utilized for tumor-specific targeting and immune escape. However, the labor-intensive purification procedures for rich-dose and high-purity homogeneous exosomes without using targeting ligands are still a challenging task. Herein, we developed a nanovector Exo-PMA/Fe-HSA@DOX through cloaked by urinary exosome membrane as a chemo/chemodynamic theranostic nano-platform for targeted homologous prostate cancer therapy which pertain to the abrogation of Epidermal Growth Factor Receptor (EGFR) and its downstream AKT/NF-kB/IkB signaling instead of ERK signaling cascades. Urinary exosomes-based nanovectors own the same urological cancer cell membrane antigen inclusive of E-cadherin, CD 47 and are free from intracellular substance such as Histone 3 and COX Ⅳ. The targeting properties of the homologous cancer cell are well preserved in Exo-PMA/Fe-HSA@DOX nanovectors in high purity. Meanwhile, the nanovectors based on urinary exosomes from prostate patients deeply penetrated into prostate cancer DU145 3D MCTS, and successfully achieve superior synergistic low-dose chemo/chemodynamic performance in vivo. In addition, the blockage of bypassing EGFR/AKT/NF-kB/IkB signaling pathway is greatly enhanced via elevated intracellular PMA/Fe-HSA@DOX nanoparticles (NPs). It is expected that the rich source and high purity of urinary exosomes offer a reliable solution for mass production of such nanovectors in the future. The targeted homologous cancer therapy based on the urinary exosomes from cancer patients exemplifies a novel targeted anticancer scheme with efficient and facile method.

P0537 - Organ-specific uptake of extracellular vesicles secreted by urological cancer cells

P0537 - Organ-specific uptake of extracellular vesicles secreted by urological cancer cells

Lopinavir-Ritonavir Combination Induces Endoplasmic Reticulum Stress and Kills Urological Cancer Cells.

Induction of endoplasmic reticulum (ER) stress is a novel approach to cancer treatment. This study investigated the ability of the clinically feasible combination of the human immunodeficiency virus protease inhibitors lopinavir and ritonavir to induce ER stress killing urological cancer cells. Renal cancer cells (769-P, 786-O) and bladder cancer cells (UMUC-3, T-24) were used to investigate the ability of the combination to induce ER stress and its mechanism of action. The combination inhibited the growth of both renal and bladder cancer cells synergistically by inducing ER stress. The combination-induced ER stress increased the expression of AMP-activated protein kinase and suppressed the mammalian target of rapamycin pathway. It also increased the expression of a tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) receptor and thereby sensitized the cancer cells to TRAIL. The combination of lopinavir and ritonavir acts against urological cancer cells by inducing ER stress synergistically.

A new niclosamide derivatives-B17 can inhibit urological cancers growth through apoptosis-related pathway.

The incidence and mortality rate of urological cancers is increasing yearly. Niclosamide has been repurposed as an anti‐cancer drug in recent years. Synthesized derivative of niclosamide was testified for its anti‐cancer activity in urological cancers. MTT assay was used to measure the cytotoxicity effect of niclosamide and its derivatives in urological cancer cell lines. Migratory ability was monitored by scratch migration assay. Apoptosis and cell cycle changes were analyzed by annexin V and PI staining. The apoptosis‐related signal proteins were evaluated by western blotting. T24 had the best drug sensitivity with the lowest IC 50 in niclosamide and B17 treatment than DU145 and Caki‐1 cells. After niclosamide and B17 treatment, the mitotic cells were decreased, but apoptotic bodies and morphology changes were not prominent in T24, Caki‐1, and DU145 cells. The migratory ability was inhibited in niclosamide treatment than control group on Caki‐1 cells and niclosamide and B17 treatment than control group on DU145 cells. Early apoptosis cells were increased after niclosamide and B17 treatment than control group without cell cycle changes in T24, Caki‐1, and DU145 cells. Programmed cell death was activated majorly through PAPR and bcl‐2 in T24 and caspase‐3 in Caki‐1 cells, respectively. Niclosamide and B17 derivative had good ability in inhibition proliferation and migratory ability in T24, Caki‐1, and DU145 cells without prominent morphology and apoptotic body changes. UCC cells are more sensitive to niclosamide and B17 treatment. Early apoptosis was induced after niclosamide and B17 treatment through different mechanisms in T24, Caki‐1, and DU145 cells.

A novel function of NUCB2 in promoting the development and invasion of renal cell carcinoma.

Previous studies have assessed nucleobindin 2 (NUCB2) expression in multiple urological cancer cell lines and detected its effect on renal cancer cell apoptosis. Additionally, certain reports have indicated a novel function of NUCB2 in promoting invasion in renal cancer. The levels of NUCB2 expression in different tumor cell lines were detected by quantitative reverse transcription polymerase chain reaction (RT-qPCR). Human NUCB2 and β-actin (ACTB) cDNA plasmids were inserted into lentivirus plasmids, which were then transfected into 786-O cells. Western blotting and RT-qPCR were then performed to determine the gene expression in NUCB2-knocked down cells. Apoptosis was also examined by flow cytometry subsequent to successful transfection. Finally, a transwell invasion assay was performed to investigate the effects on invasive abilities in renal cancer cells. The RT-qPCR results demonstrated a high expression of NUCB2 in 786-O, ACHN and LNCaP cells, and there was particularly high expression in renal cancer 786-O cells. Following successful transfection, downregulation of NUCB2 facilitated renal carcinoma cell apoptosis, as demonstrated by an increased apoptosis rate in the lenti-NUCB2-KD 786-O cells (13.72±0.84 vs. 3.32±0.10; lenti-NUCB2-KD group vs. negative control). Notably, a significant decreased invasion rate was observed in the NUCB2 knocked-down cells compared with negative control, suggesting an invasion-promoting effect of NUCB2. These results suggested a novel function of NUCB2 in the process of development and invasion in renal cell carcinoma. NUCB2 may be an important prognostic factor and target in the diagnosis and treatment of human renal cancer.

Evaluation of newly developed histone deacetylase inhibitors for the treatment of solid tumors

The inhibition of histone deacetylases (HDACs) with small‐molecule inhibitors is a new and promising approach for innovative cancer treatment. Histones are of importance for the organization of DNA in the form of chromatin. Transcriptional regulation comprises of a complex machinery of histone modifications, DNA‐methylation as well as acetylation and deacetylation of the chromatin. Dysregulation of acetylation and deacetylation of histones plays an important role in the development and progression of cancer and is often associated with an overexpression or upregulated activity of HDACs. HDAC inhibitors (HDACi) are highly versatile, and have effects on cell cycle, DNA‐repair, DNA‐damage, mitosis, apoptosis and angiogenesis. Here we investigated the suitability of newly synthesized HDACi with enhanced antitumor efficacy as compared to the already established HDACi, such as SAHA.Studies on a set of different cancer cell models (urological cancer cells with or without cisplatin resistance, neuroendocrine gastrointestinal tumor cells and esophageal cancer cells) revealed pronounced growth inhibitory and apoptosis‐inducing effects – starting at nanomolar concentrations (180nM – 1.6µM). Moreover, in vivo tests using the CAM assay method revealed antiangiogenic effects and a concomitant reduction of tumor growth of inoculated urological tumors. Further investigations will focus on the underlying mode of action of the new HDACi in terms of apoptosis, attenuation of subtype‐specific HDAC‐activity and cell cycle regulation.

Does Wnt/β-catenin pathway contribute to the stability of DNMT1 expression in urological cancer cell lines?

DNA methylation is considered as one of the most important epigenetic mechanisms and it is catalyzed by DNA methyltransferases (DNMTs). DNMT1 abundance has been frequently seen in urogenital system tumors but the reasons for this abundance are not well understood. We aimed to look into the effects of Wnt/β-catenin signaling pathway on overexpression of DNMT1 and aberrant expression of UHRF1 and HAUSP which are responsible for stability of DNMT1 at transcriptional and protein levels in urogenital cancers. In this context, firstly, Wnt/β-catenin signaling pathway was activated by using SB216763 which is a glycogen synthase kinase-3 (GSK3) β inhibitor. Cell proliferation levels in bladder cancer cells, renal cell carcinoma, and prostate cancer cells treated with GSK3β inhibitor (SB216763) were detected by WST-1 reagent. WIF-1 gene methylation profile was determined by methylation-specific PCR (MSP); expression levels of target genes β-catenin and WIF-1 by real-time PCR; and protein levels of β-catenin, DNMT1, pGSK3β(Ser9), HAUSP, and UHRF1 by Western Blot. Our results indicated that treatment with SB216763 caused an increased cell proliferation at low dose. mRNA levels of β-catenin increased after treatment with SB216273 and protein levels of pGSK3β(Ser9), β-catenin, and DNMT1 increased in comparison to control. HAUSP and UHRF1 were either up-regulated or down-regulated at the same doses depending on the type of cancer. Also, we showed that protein levels of DNMT1, β-catenin, HAUSP, and UHRF1 decreased after re-expression of WIF-1 following treatment with DAC. In Caki-2 cells, β-catenin pathway might have accounted for the stability of DNMT1 expression, whereas such relation is not valid for T24 and PC3 cells. Our findings may offer a new approach for determination of molecular effects of Wnt/β-catenin signal pathway on DNMT1. This may allow us to identify new molecular targets for the treatment of urogenital cancers.

1312 STIMULATION OF MACROPHAGE INHIBITORY CYTOKINE-1 BY PALMITIC ACID ACTIVATES PROSTATE CANCER CELL INVASIVENESS

You have accessJournal of UrologyProstate Cancer: Basic Research II1 Apr 20121312 STIMULATION OF MACROPHAGE INHIBITORY CYTOKINE-1 BY PALMITIC ACID ACTIVATES PROSTATE CANCER CELL INVASIVENESS Huang Mingguo, Shintaro Narita, Shinya Maita, Kazuyuki Numakura, Takashi Obara, Takamitsu Inoue, Yohei Horikawa, Shigeru Satoh, and Tomonori Habuchi Huang MingguoHuang Mingguo Akita, Japan More articles by this author , Shintaro NaritaShintaro Narita Akita, Japan More articles by this author , Shinya MaitaShinya Maita Akita, Japan More articles by this author , Kazuyuki NumakuraKazuyuki Numakura Akita, Japan More articles by this author , Takashi ObaraTakashi Obara Akita, Japan More articles by this author , Takamitsu InoueTakamitsu Inoue Akita, Japan More articles by this author , Yohei HorikawaYohei Horikawa Akita, Japan More articles by this author , Shigeru SatohShigeru Satoh Akita, Japan More articles by this author , and Tomonori HabuchiTomonori Habuchi Akita, Japan More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2012.02.1659AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Eating habits play important roles in prostate cancer (PCa) progression. Although lipid metabolism is strongly associated with diet-induced PCa progression, the type of fatty acid that has the most significant influence on PCa progression remains unknown. We investigate the role of long-chain fatty acids and their associated signals on PCa cell progression in vitro and in vivo. METHODS The impact of long-chain fatty acids, including palmitic acid (PA), oleic acid (OA), and linoleic acid (LA), on the proliferation and invasion of three PCa cells was assessed by cell proliferation and Matrigel invasion assays. Following stimulation by PA, the secretion and expression of transforming growth factor-beta family genes and proteins in PCa cells, other urological cancer cells, and benign prostate cells were measured using qRT-PCR, western blotting, and ELISA. The association between tumor volume and macrophage inhibitory cytokine-1 (MIC-1) expression in mouse xenograft fed with high-fat diet (HFD) or low-fat diet (LFD) was investigated. RESULTS There were no significant differences in cell proliferation in the three PCa cells treated with PA, OA, LA, and control. The three long-chain fatty acids significantly increased the invasive capacity of PC-3 and DU145 cells compared with control and the mean numbers of invaded cells were highest in PC-3 and DU145 cells treated with PA (p < 0.01 and p < 0.01, respectively). Among various cytokines, PA stimulated MIC-1 expression in the culture medium and cell extracts of three PCa cells, whereas there was no significant difference in benign prostate cells and other urological cancer cells. Furthermore, the invasive capacity of PC-3 and DU145 cells were significantly increased by treatment with recombinant MIC-1. The mean tumor volume in vivo in the HFD group was significantly higher than that in the LFD group (2569.9 vs 2010.3 mm3, p = 0.025). The mean level of serum MIC-1 in the HFD group was significantly higher than that in the LFD group (24155.2 vs 18343.8 pg/mL, p = 0.028). CONCLUSIONS Long-chain fatty acids, particularly PA, influence PCa cell invasiveness. Stimulation of MIC-1 by PA may be a significant underlying mechanism of diet-induced PCa progression. © 2012 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 187Issue 4SApril 2012Page: e532 Advertisement Copyright & Permissions© 2012 by American Urological Association Education and Research, Inc.MetricsAuthor Information Huang Mingguo Akita, Japan More articles by this author Shintaro Narita Akita, Japan More articles by this author Shinya Maita Akita, Japan More articles by this author Kazuyuki Numakura Akita, Japan More articles by this author Takashi Obara Akita, Japan More articles by this author Takamitsu Inoue Akita, Japan More articles by this author Yohei Horikawa Akita, Japan More articles by this author Shigeru Satoh Akita, Japan More articles by this author Tomonori Habuchi Akita, Japan More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Epigenetic regulation of Wnt signaling pathway in urological cancer

Constitutive activation of the Wnt signaling pathway is a common feature of solid tumors and contributes to uncontrolled cell-growth and impaired differentiation. We hypothesized that gene silencing mediated through aberrant promoter methylation of upstream Wnt antagonist genes might result in β-catenin accumulation, resulting in constitutive Wnt activation. Wnt antagonist genes (SFRP1, WIF1, APC and CDH1) and CTNNB1 promoter methylation was examined in genomic DNA extracted from 12 urological cancer cell lines and correlated with CTNNB1 mRNA expression. Promoter methylation status was then assessed in 36 BCa, 30 PCa, 31 RCT, and normal bladder mucosa (15), prostate (10) and renal (5) tissue samples. Finally, CTNNB1 mRNA relative expression levels were correlated with Wnt antagonist gene methylation status in RCT. Methylation was found in at least one Wnt antagonist gene and the CTNNB1 promoter was unmethylated in all cancer cell lines tested. When gene methylation levels were compared between cancer cell lines with high and low CTNNB1 mRNA expression, a trend was found for increased CDH1 promoter methylation levels in the former. BCa and PCa tumors demonstrated high frequency of promoter methylation at all tested genes. In RCT, CTNNB1 was unmethylated in all cases and the overall frequency of promoter methylation at the remainder genes was lower. Interestingly, median CTNNB1 mRNA expression levels were significantly higher in RCTs methylated in at least one Wnt antagonist gene promoter. We concluded that epigenetic deregulation of Wnt pathway inhibitors may contribute to aberrant activation of Wnt signaling pathway in bladder, prostate and renal tumors.

Cysteinyl-leukotriene1 receptor antagonist prevents urological cancer cell growth through early apoptosis

Recent studies have demonstrated that the cysteinyl-leukotriene1 receptor (CysLT1R) antagonist induces the growth arrest of cancer cells through apoptosis. In this study, we examined the effects of the CysLT1R antagonist on cell proliferation in urological cancer cell lines, including renal cell carcinoma, bladder cancer, prostate cancer and testicular cancer cells. The inhibitory effect of the CysLT1R antagonist on the urological cancer cells was investigated using the MTT assay and flow cytometry. The CysLT1R antagonist induced a reduction in cell viability with a half-maximal concentration of growth inhibition in all the urological cancer cell lines, and arrested the growth of the cells through early apoptosis. In conclusion, the CysLT1R antagonist may mediate potent anti-proliferative effects against urological cancer cells through early apoptosis, and may therefore serve as a novel therapeutic target in the treatment of urological cancer.

Cysteinyl-leukotriene1 receptor is a potent target for the prevention and treatment of human urological cancer.

Leukotrienes (LTs) are biologically active fatty acids derived from the oxidative metabolism of arachidonic acid through the 5-lipoxygenase pathway. LTs work to contract airway smooth muscle, increase vascular permeability and mucus secretions, and attract and activate inflammatory cells in the airways of patients with asthma. Recently, it was reported that the LTD4 receptor (cysteinylLT1 receptor; CysLT1R) plays an important role in carcinogenesis. In this study, CysLT1R expression was examined in human urological cancer cell lines (renal cell carcinoma, bladder cancer, prostate cancer and testicular cancer) using immunohistochemistry and RT-PCR. The effect of CysLT1R antagonist on these cells was also examined using the MTT assay, Hoechst staining and flow cytometry. CysLT1R expression was significantly more extensive and intense in the malignant tissues than in normal tissues. Furthermore, CysLT1R antagonist induced a reduction in malignant cell viability through early apoptosis. These results demonstrate that CysLT1R expressed in urological cancer may play a crucial role in carcinogenesis. CysLT1R may therefore be a novel target in the treatment of urological cancer.

Cysteinyl-leukotriene1 receptor antagonist prevents urological cancer cell growth through early apoptosis

Recent studies have demonstrated that the cysteinyl-leukotriene1 receptor (CysLT1R) antagonist induces the growth arrest of cancer cells through apoptosis. In this study, we examined the effects of the CysLT1R antagonist on cell proliferation in urological cancer cell lines, including renal cell carcinoma, bladder cancer, prostate cancer and testicular cancer cells. The inhibitory effect of the CysLT1R antagonist on the urological cancer cells was investigated using the MTT assay and flow cytometry. The CysLT1R antagonist induced a reduction in cell viability with a half-maximal concentration of growth inhibition in all the urological cancer cell lines, and arrested the growth of the cells through early apoptosis. In conclusion, the CysLT1R antagonist may mediate potent anti-proliferative effects against urological cancer cells through early apoptosis, and may therefore serve as a novel therapeutic target in the treatment of urological cancer.

Telmisartan inhibits human urological cancer cell growth through early apoptosis

Angiotensin II receptor blockers (ARBs) are widely used as hypertensive therapeutic agents. In addition, studies have provided evidence that ARBs have the potential to inhibit the growth of several types of cancer cells. It was reported that telmisartan (a type of ARB) has peroxisome proliferator-activated receptor (PPAR)-γ activation activity. We previously reported that the PPAR-γ ligand induces growth arrest in human urological cancer cells through apoptosis. In this study, we evaluated the effects of telmisartan and other ARBs on cell proliferation in renal cell carcinoma (RCC), bladder cancer (BC), prostate cancer (PC) and testicular cancer (TC) cell lines. The inhibitory effects of telmisartan and other ARBs (candesartan, valsartan, irbesartan and losartan) on the growth of the RCC, BC, PC and TC cell lines was investigated using an MTT assay. Flow cytometry and Hoechst staining were used to determine whether the ARBs induced apoptosis. Telmisartan caused marked growth inhibition in the urological cancer cells in a dose- and time-dependent manner. Urological cancer cells treated with 100 μM telmisartan underwent early apoptosis and DNA fragmentation. However, the other ARBs had no effect on cell proliferation in any of the urological cancer cell lines. Telmisartan may mediate potent anti-proliferative effects in urological cancer cells through PPAR-γ. Thus, telmisartan is a potent target for the prevention and treatment of human urological cancer.

A Novel Approach to Anticancer Therapies: Peroxisome Proliferator Activator-Receptor-&amp;#947; as a New Target Therapy in the Treatment of Human Urological Cancer

Peroxisome proliferator activator-receptor (PPAR)-gamma is a ligand-activated transcriptional factor belonging to a steroid receptor superfamily. PPAR-gamma plays a role in both adipocyte differentiation and carcinogenesis. Up-date, PPAR-gamma is expressed in various cancer tissues, and PPAR-gamma ligand induces growth arrest of these cancer cells. In this study, we examined the expression of PPAR-gamma in human urological cancer (including renal cell carcinoma, bladder tumor, prostate cancer and testicular cancer) by RT-PCR and immunohistochemistry, and we also examined the effect of PPAR-gamma ligand in these cells by MTT assay, flow cytometry and hoechest staining. PPAR-gamma expression was significantly more extensive and intense in malignant tissues than in normal tissues. PPAR-gamma ligand induced the reduction of malignant cell viability through early apoptosis. These results demonstrated that generated PPAR-gamma in urological cancer cells may play an important role in carcinogensis and become a new target therapy in the treatment of urological cancer.

Growing role of CD40 ligand gene transfer therapy in the management of systemic malignancies besides hepatocellular carcinomas

The article “Cationic liposome-mediated transfection of CD40 ligand gene inhibits hepatic tumor growth of hepatocellular carcinoma in mice” [doi:10.1631/jzus.B0820178] by Jiang et al.(2009) in a recent issue of the Journal of Zhejiang University SCIENCE B was highly thought provoking. The authors have clearly demonstrated the efficacy of CD40 ligand gene therapy in inhibiting the growth of hepatocellular carcinomas. The findings of Jiang et al.(2009) are highly important as they further support and corroborate the rapidly expanding role of CD40 ligand gene therapy in the management of systemic malignancies besides hepatocellular carcinomas. For instance, CD40 ligand gene transfer into cutaneous dendritic cells attenuates tumor growth in dermatological malignancies such as squamous cell carcinoma cell lines (Tomihara et al., 2008). Similar attenuation of tumor growth in gastrointestinal tumors, such as pancreatic carcinomas, has been achieved following CD40 ligand gene therapy (Serba et al., 2008; Kuhlmann et al., 2008). CD40 ligand gene transfection by using adenoviruses has also been successfully used for regressing tumor growth in pulmonary carcinomas (Wu et al., 2007). CD40 ligand gene transfection has also been used in controlling tumor growth in other systemic malignancies. For instance, urological cancer cell lines, such as the TRAMP-C2 (prostate carcinoma) cell line, show attenuated tumor growth and size following CD40 ligand gene therapy (Dzojic et al., 2006). The above examples have clearly highlighted the importance of the findings of Jiang et al.(2009). Clearly, CD40 ligand gene therapy has a major role to play in the control and management of systemic malignancies and may prove to be a major tool in medicine.

Novel HER2 selective tyrosine kinase inhibitor, TAK‐165, inhibits bladder, kidney and androgen‐independent prostate cancer in vitro and in vivo

TAK-165 is a new potent inhibitor of human epidermal growth factor receptor 2 (HER2) tyrosine kinase. Several reports suggest HER2 expression in bladder cancer, renal cell carcinoma (RCC) and androgen-independent prostate cancer. We therefore investigated the antitumor effect of TAK-165 on these urological cancer cells. Western blot analysis was performed to confirm HER2 expression in cell lines. To study in vitro efficacy, cells were treated with TAK-165 at various concentrations for 72 h and then counted using a hemocytometer. Then the IC50 value was calculated. In the xenograft model, after the tumor reached 200-300 mm3 in volume, mice were orally administered TAK-165 10 mg/kg per day or 20 mg/kg per day or saline for 14 consecutive days (n=6-8). HER2 expression was observed in HT1376, UMUC3, T24 (bladder), ACHN (kidney), DU145, LNCaP, LN-REC4 (prostate), although the expression level in these cells was weak compared with BT474 (a breast cancer cell line which expresses HER2 strongly). IC50 was varied from 0.09 to greater than 25 micromol/L in the bladder cancer cell line. ACHN cells were less sensitive in vitro. The prostate cancer cell lines studied were all sensitive (IC50 0.053-4.62 micromol/L). In the xenograft model, treatment with TAK-165 significantly inhibited growth of UMUC-3, ACHN, and LN-REC4. The antitumor effect (T/C [%]=growth of TAK-165 treated tumor/average growth of control tumorx100) after 14 days treatment were 22.9%, 26.0%, and 26.5% in UMUC3, ACHN and LN-REC4, respectively. TAK-165 may be a hopeful new agent for bladder, kidney and androgen-independent prostate cancer.