scholarly journals Knockdown of RRM1 with Adenoviral shRNA Vectors to Inhibit Tumor Cell Viability and Increase Chemotherapeutic Sensitivity to Gemcitabine in Bladder Cancer Cells

2021 ◽  
Vol 22 (8) ◽  
pp. 4102
Author(s):  
Xia Zhang ◽  
Rikiya Taoka ◽  
Dage Liu ◽  
Yuki Matsuoka ◽  
Yoichiro Tohi ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Bladder Cancer ◽  
Cancer Cells ◽  
Cell Lines ◽  
Cancer Cell ◽  
Bladder Cancer Cell ◽  
Human Tumor Xenograft ◽  
Antitumor Effects ◽  
Chemotherapeutic Sensitivity ◽  
Bladder Cancer Cells

RRM1—an important DNA replication/repair enzyme—is the primary molecular gemcitabine (GEM) target. High RRM1-expression associates with gemcitabine-resistance in various cancers and RRM1 inhibition may provide novel cancer treatment approaches. Our study elucidates how RRM1 inhibition affects cancer cell proliferation and influences gemcitabine-resistant bladder cancer cells. Of nine bladder cancer cell lines investigated, two RRM1 highly expressed cells, 253J and RT112, were selected for further experimentation. An RRM1-targeting shRNA was cloned into adenoviral vector, Ad-shRRM1. Gene and protein expression were investigated using real-time PCR and western blotting. Cell proliferation rate and chemotherapeutic sensitivity to GEM were assessed by MTT assay. A human tumor xenograft model was prepared by implanting RRM1 highly expressed tumors, derived from RT112 cells, in nude mice. Infection with Ad-shRRM1 effectively downregulated RRM1 expression, significantly inhibiting cell growth in both RRM1 highly expressed tumor cells. In vivo, Ad-shRRM1 treatment had pronounced antitumor effects against RRM1 highly expressed tumor xenografts (p < 0.05). Moreover, combination of Ad-shRRM1 and GEM inhibited cell proliferation in both cell lines significantly more than either treatment individually. Cancer gene therapy using anti-RRM1 shRNA has pronounced antitumor effects against RRM1 highly expressed tumors, and RRM1 inhibition specifically increases bladder cancer cell GEM-sensitivity. Ad-shRRM1/GEM combination therapy may offer new treatment options for patients with GEM-resistant bladder tumors.

scholarly journals TRAF4 inhibits bladder cancer progression by promoting BMP/SMAD signalling pathway

2020 ◽  
Author(s):  
Prasanna Vasudevan Iyengar ◽  
Dieuwke Louise Marvin ◽  
Dilraj Lama ◽  
Tuan Zea Tan ◽  
Sudha Suriyamurthy ◽  
...  
Keyword(s):  
Bladder Cancer ◽  
Cancer Cells ◽  
Cell Lines ◽  
Cancer Cell ◽  
Ubiquitin Ligase ◽  
E3 Ubiquitin Ligase ◽  
Cancer Cell Lines ◽  
Bladder Cancer Cell ◽  
Over Expression ◽  
Bladder Cancer Cells

AbstractBladder cancer is one of the most prevalent cancer types in the world, frequently exhibiting invasion and metastasis and therefore associated with poor prognosis. It is a progressive disease with high recurrence rates even after surgery, which calls for the urgent need for early intervention and diagnosis. The E3 ubiquitin ligase TNF Receptor Associated Factor 4 (TRAF4) has been largely implicated as a tumour-promoting element in a wide range of cancers. Over-expression and amplification of this gene product has been a common observation in breast and other metastatic tumours. In contrast, we observed that expression of TRAF4 negatively correlated with overall patient survival. Moreover, its expression was repressed at epigenetic levels in aggressive bladder cancer cells. We also describe an ERK kinase phosphorylation site on TRAF4 that inhibits its stability and localization to plasma membrane in such cells. Furthermore, knockdown of TRAF4 in epithelial bladder cancer cell lines leads to gain of mesenchymal genes and a loss of epithelial integrity. Reciprocally, stable over-expression of TRAF4 in mesenchymal cells leads to decreased migratory and invasive properties. Transcriptomic analysis upon TRAF4 mis-expression in bladder cancer cell lines revealed that higher TRAF4 expression enhanced BMP/SMAD and dampened NF-κB signalling pathways. Importantly, this notion was confirmed in bladder cancer patient material. Mechanistically, we showed that TRAF4 targets the E3 ubiquitin ligase SMURF1, a negative regulator of BMP/SMAD signalling, for proteasomal degradation in bladder cancer cells. We show that genetic and pharmacological inhibition of SMURF1 or its function inhibited migration of aggressive (mesenchymal) bladder cancer cells.

scholarly journals The lncRNA DLX6-AS1 promoted cell proliferation, invasion, migration and epithelial-to-mesenchymal transition in bladder cancer via modulating Wnt/β-catenin signaling pathway

2019 ◽  
Vol 19 (1) ◽  
Author(s):  
Jinan Guo ◽  
Zhixin Chen ◽  
Hongtao Jiang ◽  
Zhou Yu ◽  
Junming Peng ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Bladder Cancer ◽  
Cancer Cells ◽  
Cancer Cell ◽  
Cancer Progression ◽  
Bladder Cancer Cell ◽  
Mesenchymal Transition ◽  
Bladder Cancer Cells

Abstract Background Bladder cancer is the most common human urological malignancies with poor prognosis, and the pathophysiology of bladder cancer involves multi-linkages of regulatory networks in the bladder cancer cells. Recently, the long noncoding RNAs (lncRNAs) have been extensively studied for their role on bladder cancer progression. In this study, we evaluated the expression of DLX6 Antisense RNA 1 (DLX6-AS1) in the cancerous bladder tissues and studied the possible mechanisms of DLX6-AS1 in regulating bladder cancer progression. Methods Gene expression was determined by qRT-PCR; protein expression levels were evaluated by western blot assay; in vitro functional assays were used to determine cell proliferation, invasion and migration; nude mice were used to establish the tumor xenograft model. Results Our results showed the up-regulation of DLX6-AS1 in cancerous bladder cancer tissues and bladder cell lines, and high expression of DLX6-AS1 was correlated with advance TNM stage, lymphatic node metastasis and distant metastasis. The in vitro experimental data showed that DLX6-AS1 overexpression promoted bladder cancer cell growth, proliferation, invasion, migration and epithelial-to-mesenchymal transition (EMT); while DLX6-AS1 inhibition exerted tumor suppressive actions on bladder cancer cells. Further results showed that DLX6-AS1 overexpression increased the activity of Wnt/β-catenin signaling, and the oncogenic role of DLX6-AS1 in bladder cancer cells was abolished by the presence of XAV939. On the other hand, DLX6-AS1 knockdown suppressed the activity of Wnt/β-catenin signaling, and the tumor-suppressive effects of DLX6-AS1 knockdown partially attenuated by lithium chloride and SB-216763 pretreatment. The in vivo tumor growth study showed that DLX6-AS1 knockdown suppressed tumor growth of T24 cells and suppressed EMT and Wnt/β-catenin signaling in the tumor tissues. Conclusion Collectively, the present study for the first time identified the up-regulation of DLX6-AS1 in clinical bladder cancer tissues and in bladder cancer cell lines. The results from in vitro and in vivo assays implied that DLX6-AS1 exerted enhanced effects on bladder cancer cell proliferation, invasion and migration partly via modulating EMT and the activity of Wnt/β-catenin signaling pathway.

scholarly journals Chloride Ion Transport Blockade Enhances Cytocidal Effects of Sterile Water on Bladder Cancer Cells

2021 ◽  
Author(s):  
Yuki Matsuoka ◽  
Rikiya Taoka ◽  
Yoichiro Tohi ◽  
Zhang Xia ◽  
Mikio Sugimoto
Keyword(s):  
Bladder Cancer ◽  
Cancer Cells ◽  
Cell Lines ◽  
Cancer Cell ◽  
Channel Blocker ◽  
Bladder Cancer Cell ◽  
Channel Blockers ◽  
Sterile Water ◽  
Hypotonic Shock ◽  
Bladder Cancer Cells

Abstract Background:Bladder cancer is a major health concern worldwide. The conventional intravesical Bacillus Calmette–Guérin therapy has certain shortcomings; thereby, demanding novel alternatives. Although sterile water is a probable agent for such novel intravesical therapies, bladder cancer cell lines differ in their sensitivity to hypotonic shock due to sterile water. Therefore, we aimed to investigate whether Cl- channel blockers enhance the cytocidal effect of hypotonic shock on bladder cancer cells resistant to sterile water.Methods:Bladder cancer cell lines of varying grades (RT112, T24, and J82) were exposed to sterile water, and morphological changes were closely observed using microscopy. Sterile water-induced changes in cell membrane integrity and cell viability were analyzed to determine the effects of hypotonic shock. These effects were further analyzed using a Cl- channel blocker.Results:T24 and J82 cells started swelling immediately upon exposure to sterile water and ruptured within 10 min. RT112 cells demonstrated limited hypotonic swelling with few cell ruptures. After treatment with the Cl- channel blocker, RT112 cells ruptured faster as compared to that in cells treated with sterile water. The percentages of viable dimethylsulfoxide and 5-nitro-2-(3-phenylpropylamino) benzoic acid -treated (50, 100, 200, and 300 µM) RT112 cells after 10 min of exposure to sterile water were 13.6 % ± 3.4 %, 6.3 % ± 1.2 %, 2.0 % ± 1.1 %, 0.7 % ± 0.7 %, and 0 %, respectively.Conclusions:Taken together, the Cl- channel blockers enhanced the cytocidal effects of hypotonic shock in bladder cancer cells. Intravesical therapy with sterile water after treatment with a Cl- channel blocker represents a potential new adjuvant therapy after TURBT with high efficacy.

Cancer-specific nanotheranostics to improve the diagnosis and treatment of bladder cancer.

2015 ◽  
Vol 33 (7_suppl) ◽  
pp. 323-323
Author(s):  
Chong-Xian Pan ◽  
Tzu-yin Lin ◽  
Hongyong Zhang ◽  
Yuanpei Li ◽  
Susan D. Airhart ◽  
...  
Keyword(s):  
Bladder Cancer ◽  
Phase I ◽  
Cancer Cells ◽  
Cell Lines ◽  
Cancer Cell ◽  
Intravesical Instillation ◽  
Bladder Cancer Cell ◽  
Photodynamic Diagnosis ◽  
Drug Load ◽  
Bladder Cancer Cells

323 Background: We recently developed a bladder cancer-specific targeting ligand named PLZ4, nanometer-scale micelles and nanoporphyrin. Here we report the diagnostic and therapeutic applications of these nanotheranostics coated with PLZ4. Methods: PLZ4 was synthesized through solid phase synthesis. Bladder cancer-specific PLZ4-coated nanomicelles (PNM) and nanoporphyrin (PNP) were developed through conjugating the nanotheranostics with PLZ4 on the surface, and loaded with therapeutic and/or imaging agents in the core. Bladder cancer cell lines and patient-derived xenografts (PDXs) were used to determine the diagnostic and therapeutic applications. Results: In vitro studies with cell lines revealed that both PNM and PNP could specifically deliver the drug load to bladder cancer cells, but not to adjacent confounding cells. After intravenous injection, PNM loaded with paclitaxel could specifically deliver the drug load to xenografts developed from a human and a dog bladder cancer cell line, and a PDX, but not to lung cancer xenografts in the same mice. These paclitaxel-loaded PNM could overcome cisplatin resistance, and prolong the overall survival of mice carrying PDXs from 27 days with free paclitaxel to 76 days (p<0.0001). PNP can be used for photodynamic diagnosis and therapy while being able to chelate gadolinium for cancer-specific magnetic resonance imaging (MRI), and load chemotherapeutic drugs for cancer-specific targeted chemotherapy. It is over 50 times more potent that 5-aminolevulinic acid in photodynamic therapy (p<0.0001). After intravesical instillation into the bladder cavity of an orthotopic PDX model, PNP could specifically target bladder cancer cells, but not adjacent normal urothelial cells in the same bladder. Conclusions: PNM and PNP can potentially be used for diagnosis, imaging detection and cancer-specific targeted delivery of therapeutic agents of both non-myoinvasive and advanced bladder cancer. A phase I clinical trial of PNM for intravesical instillation in human patients with non-myoinvasive bladder cancer, and another phase I trial with PNP for photodynamic diagnosis and therapy in dog bladder cancer patients have been funded.

scholarly journals miR-200a-3p Facilitates Bladder Cancer Cell Proliferation by Targeting A20

2021 ◽  
Author(s):  
pei wan ◽  
Zhilin Chen ◽  
Minzhi Huang ◽  
Huiming Jiang ◽  
Huajun Wu ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Bladder Cancer ◽  
Cancer Cells ◽  
Cancer Cell ◽  
Bladder Cancer Cell ◽  
Cancer Cell Proliferation ◽  
Human Bladder Cancer ◽  
Human Bladder ◽  
Bladder Cancer Cells ◽  
Cancer Tissues

Abstract BackgroundMicroRNAs (miRs) are endogenous, single-stranded, non-coding RNAs that are involved in various physiological processes, development and the progression of various types of cancer. The role of miR-200a-3p in various types of cancer has been previously reported. The present study aimed to investigate the expression levels of miR-200a-3p in human bladder cancer, as well as its potential role in disease pathogenesis.MethodsIn vitro: Agilent miRNA microarray and QPCR analysis of miR-200a-3p expression in bladder cancer. Gene overexpression and interference technology analysis of the effects of miR-200a-3p and de-ubiquituination enzyme TNFα induced protein 3 (A20) on the function of bladder cancer cells; dual luciferase technology for promoter efficiency analysis; Animal experiments: Nude mice are used for tumor formation experiments, and the effects of genes on tumors are directly analyzed on animals;ResultsCompared with adjacent non-tumor tissues, miR-200a-3p expression levels were significantly upregulated in bladder cancer tissues. Moreover, increased miR-200a-3p expression was significantly associated with distant metastasis and advanced stage. In addition, compared with the miR-Negative control (NC) group, miR-200a-3p overexpression promoted bladder cancer cell proliferation, migration, invasion, cell cycle and release of inflammatory cytokines, but inhibited cell apoptosis. Mechanistically, de-ubiquituination enzyme TNFα induced protein 3 (A20) was identified as a target gene of miR-200a-3p in bladder cancer cell lines. An inverse association between miR-200a-3p expression and A20 expression in bladder cancer tissues and cell lines was also identified. A20 overexpression in miR-200a-3p-overexpression bladder cancer cells attenuated miR-200a-3p overexpression-mediated effects on cell proliferation, migration, apoptosis and cytokine production. Moreover, compared with the miR-NC group, miR-200a-3p overexpression significantly promoted tumor growth in vivo, and A20 overexpression blocked the promoting effect of miR-200a-3p on bladder cancer. ConclusionsThe results of the present study indicated that miR-200a-3p might serve as an oncogene in human bladder cancer by targeting a novel gene A20; therefore, miR-200a-3p and A20 might serve as novel therapeutic targets for bladder cancer.

scholarly journals miR-489 Suppresses Proliferation and Invasion of Human Bladder Cancer Cells

2016 ◽  
Vol 24 (6) ◽  
pp. 391-398 ◽  
Author(s):  
Jing Li ◽  
Weixing Qu ◽  
Yazhou Jiang ◽  
Yi Sun ◽  
Yongyi Cheng ◽  
...  
Keyword(s):  
Bladder Cancer ◽  
Cancer Cells ◽  
Cell Lines ◽  
Cancer Cell ◽  
Bladder Cancer Cell ◽  
Human Bladder Cancer ◽  
Human Bladder ◽  
Bladder Cancer Cells ◽  
Human Bladder Cancer Cells ◽  
Proliferation And Invasion

MicroRNAs (miRNAs) have been shown to be involved in bladder cancer progression. miR-489 (also known as miR-489-3p) was recently reported to be a tumor suppressor in several cancers. However, its exact role and mechanism in the progression of bladder cancer are largely unknown. In this study, we explore the role of miR-489 in the proliferation and invasion of human bladder cancer cells. The miR-489 expression levels were detected in bladder cancer and normal adjacent tissues, as well as in human normal bladder epithelial cells and bladder cancer cell lines. The results showed that miR-489 was sharply reduced in bladder cancer tissues and cell lines. Then the miR-489 mimic or oligo anta-miR-489 was transfected into T24 and UMUC3 bladder cancer cell lines. The results showed that the miR-489 mimic greatly increased the miR-489 level and significantly decreased the proliferation and invasion of T24 and UMUC3 cells. In contrast, the anta-miR-489 had a completely opposite effect on miR-489 expression, cell proliferation, and cell invasion. Moreover, bioinformatics and luciferase reporter gene assays confirmed that miR-489 targeted the mRNA 3′-untranslated region (3′-UTR) region of Jagged1 (JAG1), a Notch ligand. In conclusion, miR-489 suppressed proliferation and invasion of human bladder cancer cells.

scholarly journals Lnc-STYK1-2 regulates bladder cancer cell proliferation, migration, and invasion by targeting miR-146b-5p expression and AKT/STAT3/NF-kB signaling

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Ranran Dai ◽  
Qingping Jiang ◽  
You Zhou ◽  
Ruifeng Lin ◽  
Hai Lin ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Bladder Cancer ◽  
Cancer Cells ◽  
Cancer Cell ◽  
Noncoding Rnas ◽  
Bladder Cancer Cell ◽  
Luciferase Reporter ◽  
Migration And Invasion ◽  
Cancer Cell Proliferation ◽  
Bladder Cancer Cells

Abstract Background Epigenetic modulation by noncoding RNAs substantially contributes to human cancer development, but noncoding RNAs involvement in bladder cancer remains poorly understood. This study investigated the role of long noncoding RNA (lncRNA) lnc-STYK1-2 in tumorigenesis in cancerous bladder cells. Methods Differential lncRNA and mRNA profiles were characterized by high-throughput RNA sequencing combined with validation via quantitative PCR. Bladder cancer cell proliferation was assessed through MTS, and bladder cancer cell migration and invasion were assessed through a Transwell system. The in vivo tumorigenesis of bladder cancer cells was evaluated using the cancer cell line-based xenograft model. The dual-luciferase reporter assay verified the association of miR-146b-5p with lnc-STYK1-2 and the target gene. Protein abundances and phosphorylation were detected by Western blotting. Results Alterations in lncRNA profiles, including decreased lnc-STYK1-2 expression, were detected in bladder cancer tissues compared with adjacent noncancerous tissues. lnc-STYK1-2 silencing effectively promoted proliferation, migration, and invasion in two bladder cancer cell lines, 5637 and T24, and their tumorigenesis in nude mice. lnc-STYK1-2 siRNA promoted miR-146b-5p and reduced ITGA2 expression in bladder cancer cells. Moreover, miR-146b-5p suppressed ITGA2 expression in bladder cancer cells through direct association. Also, lnc-STYK1-2 directly associated with miR-146b-5p. Finally, miR-146b-5p inhibitors abrogated the alterations in bladder cell functions, ITGA2 expression, and phosphorylation of AKT, STAT3, and P65 proteins in 5637 and T24 cells induced by lnc-STYK1-2 silencing. Conclusion lnc-STYK1-2 inhibited bladder cancer cell proliferation, migration, and tumorigenesis by targeting miR-146b-5p to regulate ITGA2 expression and AKT/STAT3/NF-kB signaling.

scholarly journals Down-regulation LncRNA-SNHG15 contributes to proliferation and invasion of bladder cancer cells

BMC Urology ◽  
2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Aldhabi Mokhtar ◽  
Chuize Kong ◽  
Zhe Zhang ◽  
Yan Du
Keyword(s):  
Cell Proliferation ◽  
Bladder Cancer ◽  
Cancer Cells ◽  
Cell Lines ◽  
Tumor Stage ◽  
Rt Pcr ◽  
Down Regulation ◽  
Bladder Cancer Cells ◽  
Cancer Tissues ◽  
Proliferation And Invasion

Abstract Objectives The aim of this study was to investigate the effect of lncRNA-SNHG15 in bladder carcinoma using cell lines experiments and the relationship between clinical characteristics and lncRNA-SNHG15 expression was analyzed. Methods Bladder cancer tissues and near-cancer tissues were collected. The real-time PCR (RT-PCR) was used to detect the expression of lncRNA-SNHG15 in tissues and cell lines. The expression of lncRNA-SNHG15 was downregulated by interference (siRNA), as detected by RT-PCR, that was used to determine the efficiency of the interference. CCK-8 and Transwell assays were used to evaluate the effect of lncRNA-SNHG15 on the proliferation and invasion capability of bladder cancer cells. The t-test was used for Statistical analyses, which were carried out using the Statistical Graph pad 8.0.1.224 software. Result The expression of lncRNA-SNHG15 was up regulated in 5637, UMUC3 and T24 cell lines compared with corresponding normal controls (P < 0.05). Up regulation was positively related to tumor stage (P = 0.015). And tumor size (P = 0.0465). The down-regulation of lncRNA-SNHG15 with siRNA significantly inhibited UMUC3 and T24 cell proliferation and invasion. Conclusion This study showed that lncRNA-SNHG15 is overexpressed in bladder cancer tissues and (5637, UMUC3 T24) cell lines. Up regulation was positively related to tumor stage (P = 0.015), and tumor size (P = 0.0465). Down-regulation of lncRNA-SNHG15 by siRNA significantly inhibited UMUC3 and T24 cell proliferation and invasion, indicating a potential molecular target for future tumor targeted therapy.

Apoptosis triggered by isoquercitrin in bladder cancer cells by activating the AMPK-activated protein kinase pathway

2017 ◽  
Vol 8 (10) ◽  
pp. 3707-3722 ◽  
Author(s):  
Ping Wu ◽  
Siyuan Liu ◽  
Jianyu Su ◽  
Jianping Chen ◽  
Lin Li ◽  
...  
Keyword(s):  
Bladder Cancer ◽  
Protein Kinase ◽  
Cancer Cells ◽  
Cancer Cell ◽  
Cell Metabolism ◽  
Bladder Cancer Cell ◽  
Bladder Cancer Cells ◽  
Immunoblotting Assay ◽  
Cancer Cell Metabolism ◽  
Kinase Pathway

Our findings provide comprehensive evidence that isoquercitrin (ISO) influenced T24 bladder cancer cell metabolism by activating the AMPK pathway as identified by combination with metabolomics and immunoblotting assay.

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