Human Bladder Cancer Cell T24 Research Articles

CRH upregulates supervillin through ERK and AKT pathways to promote bladder cancer cell migration.

Corticotropin-releasing hormone (CRH) has been well documented playing a role in the regulation of cellular processes, immune responses, and inflammatory processes that can influence the occurrence and development of tumors. Supervillin (SVIL) is a membrane-associated and actin-binding protein, which is actively involved in the proliferation, spread, and migration of cancer cells. This work investigated CRH's influence on bladder cancer cells'migration and relevant mechanisms. By using human bladder cancer cells T24 and RT4 in wound healing experiments and transwell assay, we found that the migration ability of the T24 cells was significantly increased after CRH treatment. In vivo experiments showed that CRH significantly promoted the metastases of T24 cells in cell line-derived xenograft (CDX) mouse model. Interestingly, downregulation of SVIL by SVIL-specifc small hairpin RNAs significantly reduced the promoting effect of CRH on bladder cancer cell migration. Furthermore, CRH significantly increased SVILmessenger RNA and protein expression in T24 cells, accompanied with AKT andERK phosphorylation in T24 cells. Pretreatment with AKT inhibitor (MK2206) blocked the CRH-induced SVIL expression and ERK phosphorylation. Also, inhibition of ERK signaling pathway by U0126 significantly reduced the CRH-induced SVIL expression and AKT phosphorylation. It suggested that cross-talking between AKT and ERK pathways was involved in the effect of CRH on SVIL. Taken together, we demonstrated that CRH induced migration of bladder cancer cells, in which AKT andERK pathways -SVIL played a key role.

New cyclic C-geranylflavanones isolated from Paulownia fortunei fruits with their anti-proliferative effects on three cancer cell lines

Seven new C-geranylated flavanones, fortunones F - L (1–7), were isolated from the fresh mature fruits of Paulownia fortunei (Seem.) Hemsl. Their structures were determined by extensive spectroscopic data interpretation (UV, IR, HRMS, NMR, and CD). These new isolated compounds were all with a cyclic side chain modified from the geranyl group. Among them, compounds 1–3 all possessed a dicyclic geranyl modification, which was described firstly for Paulownia C-geranylated flavonoids. All the isolated compounds were subjected to the cytotoxic assay on human lung cancer cell A549, mouse prostate cancer cell RM1 and human bladder cancer cell T24, respectively. Results indicated A549 cell line was more sensitive to C-geranylated flavanones than the other two cancer cell lines and compounds 1, 7 and 8 exhibited potential anti-tumor effects (IC50 ˂ 10 μM). Further research revealed the effective C-geranylated flavanones could exert their anti-proliferative activity on A549 cells by inducing apoptosis and blocking cells in G1 phase.

Farnesoid X Receptor Overexpression Decreases the Migration, Invasion and Angiogenesis of Human Bladder Cancers via AMPK Activation and Cholesterol Biosynthesis Inhibition.

Simple SummaryOur previous studies characterized that FXR overexpression results in the inhibition of migratory, adhesive and angiogenic abilities through the proteosome degradation pathway in human bladder cancer cells. Since cholesterol metabolism plays an important role during cancer progression, we investigated the role of cholesterol biosynthesis-related proteins expression and their signal transduction pathway in human bladder cancer cells. We confirmed FXR overexpression decreased muscle invasive human bladder cancer cell T24’s metastatic ability in nude mice animal models. Moreover, statin usage showed potent enough efficacy to strengthen the enhancement of FXR-inhibited migration, adhesion and angiogenesis in human urothelial carcinoma cells. Additionally, clinical data showed survival benefits of statin usage in stage 0–I bladder cancer patients. Our results suggested that FXR overexpression, combined with atorvastatin treatment, may provide a potential therapeutic strategy for the treatment of human urothelial carcinoma in the future.Bladder cancer is one of the most prevailing cancers worldwide. Although treatments for urothelial carcinoma have improved, the rate of recurrence observed in the clinic is still high. The aim of this study was to evaluate whether cholesterol biosynthesis is involved in the effect of Farnesoid X Receptor (FXR) on bladder cancers. FXR overexpression contributed to activation of 5′ AMP-activated protein kinase (AMPK) and decreased cholesterol levels. FXR overexpression reduced cholesterol biosynthesis and secretion by downregulating Sterol Regulatory Element Binding Protein 2 (SREBP2) and 3-Hydroxy-3-Methylglutaryl-CoA Reductase (HMGCR) expression. In addition, an AMPK inhibitor, dorsomorphin, reversed the inhibition of migration, invasion and angiogenesis by FXR overexpression. In a metastatic xenograft animal study, FXR overexpression suppressed bladder cancer lung metastasis by decreasing matrix metalloproteinase-2 (MMP2), SREBP2 and HMGCR expression. Moreover, FXR overexpression combined with atorvastatin treatment further enhanced the downregulation of the migratory, adhesive, invasive and angiogenic properties in human urothelial carcinoma. In clinical observations, statin administration was associated with better survival rates of early-stage bladder cancer patients. Our results may provide guidance for improving therapeutic strategies for the treatment of urothelial carcinoma.

Chinese Propolis Inhibits the Proliferation of Human Gastric Cancer Cells by Inducing Apoptosis and Cell Cycle Arrest.

Special Chinese propolis sourced from the Changbai Mountains (CBMP) in Northeast China is rich in specific flavonoids and phenolic acids and its bioactivity has not been reported. This study aimed to investigate the antiproliferative effect of CBMP on cancer cells and its molecular mechanisms. Different cancer cell lines were treated with the ethanol extracts of CBMP for 24 hours before the cell viability and mechanism measurements. The results showed CBMP had weak activities against human pancreatic cancer cell PANC1, human lung cancer cell A549, human colon cancer cell HCT116, human liver cancer cell HepG2, human bladder cancer cell T24, and human breast cancer cell MDA-MB-231, but it significantly inhibited the growth of human gastric cancer SGC-7901 cells, caused cell apoptosis and cell cycle arrest in S phase, with increased production of reactive oxygen species (ROS) and reduced mitochondrial membrane potential (MMP). The results indicate that Chinese propolis sourced from the Changbai Mountains selectively inhibits the proliferation of human gastric cancer SGC-7901 cells by inducing both death receptor-induced apoptosis and mitochondria-mediated apoptosis, and cell cycle arrest in S phase. These activities and mechanisms help understand the anticancer action of propolis and its active compounds.

Effect of ERH gene on migration and invasion of human bladder cancer cells T24 and 5637

Objective To study the effect of enhancer of rudimentary homolog (ERH) gene on migration and invasion in human bladder cancer T24 and 5637 cells. Methods After knocking out the ERH gene of human bladder cancer T24 and 5637 cells, Wound healing assay, Transwell cell migration assay and Transwell cell invasion assay were used to verify the migration and invasion function. Cell migration related protein was detected by Western blot. Nude mouse tail vein transfer assay was used to study the metastasis ability of bladder cancer cells in vivo. Results ⑴ The Wound healing assay showed that there were significant differences in the migration cell counts of human bladder cancer 5637 and T24 (P<0.05). ⑵ There were significant differences in migration and invasion cell counts of Transwell assay (P<0.05). ⑶ Western blot showed that the expression of E-Cadherin in human bladder cancer 5637 cells and T24 cells was significantly increased (P<0.05) after knocking out ERH gene, while the expression of Fibronectin, Twist, Vimentin and Snail2 protein were significantly decreased (P<0.05). ⑷ Nude mouse tail vein transfer assay showed that lung metastases were significantly reduced in the ERH knockout group compared with the normal ERH group. Conclusions Both in vitro and in vivo experiments suggest that ERH knockout affects the migration and invasion of human bladder cancer T24 and 5637 cells. Key words: Transcription factors; Cell cycle proteins; Urinary bladder neoplasms; Cell migration assays; In vitro

MiR-204 acts as a tumor suppressor in human bladder cancer cell T24 by targeting antiapoptotic BCL2

ObjectiveBladder cancer (BC) is one of the most common noncutaneous malignant cancers with high mortality and recurrence rates in the general population, particularly in men. We previously reported detecting dysregulated micro-RNAs (miRNAs) in human BC tissues. Using an miRNA targeting database, we found that miR-204, which is downregulated in BC, targets the B-cell lymphoma 2 gene (BCL2). We hypothesized that the overexpression of miR-204 induces apoptosis by repressing BCL2 expression and causing cancer cell death. Materials and methodsTo overexpress miR-204 in cancer cells, a vector that expresses miR-204 was transfected into human BC, embryonic kidney, and cervical cancer cells. The expression of miR-204 in the transfected and mock cells was monitored through miRNA quantitative polymerase chain reactions (Q-PCRs). Luciferase reporter assays were performed to verify the direct binding of miR-204 to targeted sites on the BCL2 transcripts. The BCL2 mRNA and protein expression levels in the miR-204-transfected cells were measured using Q-PCRs and Western blotting, respectively. The cell viability of the miR-204 transfected cells was detected through WST-1 assays. The induction of apoptosis was determined by the increased release of cytochrome c, cleavage caspase 3, caspase 3/7 activity, and DNA fragmentation. ResultsThe expression of miR-204 in the transfected cells was elevated to 6.2-fold, compared with the controls, and the function of matured miR-204 was confirmed through the luciferase reporter assays. The expressions of BCL2 mRNA and protein were reduced in the miR-204-transfected cells, which generated increased DNA fragmentation, caspase 3 cleavage, caspase 3/7 activity, and reduced cell viability. Cotransfection of the reporter vector that harbored the BCL2 3′-untranslated region to compete with endogenous transcripts partially rescued the apoptosis that was induced by the overexpressed miR-204. This finding further supported that miR-204 targeting BCL2 induces apoptosis in BC cells. ConclusionmiR-204 targets BCL2 in human BC cells. The overexpression of miR-204 in tumors could thus be a novel strategy for inducing cell death by targeting antiapoptotic BCL2.

Catalpol Inhibited the Proliferation of T24 Human Bladder Cancer Cells by Inducing Apoptosis Through the Blockade of Akt-Mediated Anti-apoptotic Signaling.

Catalpol is an iridoid glucoside extracted from the traditional Chinese medicinal plant called Rehmannia glutinosa, and it is reported to possess a variety of pharmacological activities. The goal of this research was to explore the effect of catalpol on the human bladder cancer cells T24. The cells were treated for different durations with various concentrations of catalpol. Then the viability, mobility, and invasiveness of the cells were determined by MTT assay and flow cytometry, respectively. Catalpol was found to significantly suppress the proliferation, migration, and invasiveness of T24 bladder cancer cells in a dose- and time-dependent manner. The results also revealed that catalpol promoted apoptosis and caused the cell-cycle arrest at G2/M phase. Furthermore, the catalpol-mediated apoptosis was found to involve the modulation of PI3K/Akt pathway and inhibit the expression of anti-apoptotic B cell lymphoma-2 (Bcl-2) family proteins. Also, the expressions of Bcl-2 proapoptotic effectors, Bcl-2-associated X protein, and Bcl-2 associated death promoter were up-regulated in the cells treated with catalpol. The enzymes involved in the ultimate process of apoptosis, active caspase-3 and poly ADP ribose polymerase were elevated, and the latter was observed to be cleaved, indicating that catalpol-accelerated cell apoptosis involved a caspase-dependent pathway. These results suggest that catalpol might serve to be a promising therapeutic agent for the treatment of human bladder cancer.

Epigallocatechin-3-gallate inducing apoptosis of human bladder cancer cell T24 and its impact on the expressions of Caspase-3 protein

Objective To investigate the effects of different concentrations of epigallocatechin-3-gallate (EGCG) on apoptosis of T24 cells and the expressions of Caspase-3 protein.Methods T24 cells were cultured in vitro,and incubated with different concentrations of EGCG (0,20,40,80 mg/L,respectively).The cell proliferation was measured by growth rate analysis and cloning forming assay.The apoptosis of T24 cells was tested by flow cytometry.The expression of Caspase-3 was detected by using reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting.Results EGCG inhibited the proliferation of T24 cells.After T24 cells were stimulated with different concentrations of EGCG (0,20,40,80 mg/L),the cloning number of cells was 200 ± 43,120 ± 32,65 ± 24 and 35 ± 13 respectively ( P ＜ 0.05 ).Proliferation inhibition and apoptosis rate was increased with the increased of EGCG concentration.EGCG up-regulated the expression of caspase-3 in T24 cells in a concentration-dependent manner.Conclusion EGCG can inhibit the growth of T24 cells and induce apoptosis probably by up-regulating the expression of caspase-3. Key words: Bladder cancer; Epigallocatechin-3-gallate; Apoptosis