phenanthrenes and Urinary-Bladder-Neoplasms

The phosphorylation of pyruvate dehydrogenase (PDH) by pyruvate dehydrogenase kinase (PDK) 4 inhibits its ability to induce a glycolytic shift. PDK4 expression is upregulated in various types of human cancer. Because PDK4 regulation is critical for metabolic changes in cancer cells, it is an attractive target for cancer therapy given its ability to shift glucose metabolism. It was previously shown that a novel PDK4 inhibitor, cryptotanshinone (CPT), suppressed the three‑dimensional (3D)‑spheroid formation of pancreatic and colorectal cancer cells. In the present study, the effects of CPT on the invasiveness of bladder cancer cells were investigated. CPT significantly suppressed the invasiveness and 3D‑spheroid formation of T24 and J82 bladder cancer cells. CPT also suppressed the phosphorylation of PDH and β‑catenin, as well as the expression of N‑cadherin, which are all critical for inducing epithelial‑mesenchymal transition (EMT). The knockdown of β‑catenin or PDK4 using specific small interfering RNAs suppressed N‑cadherin expression and invasiveness in T24 cells. An mTOR inhibitor also suppressed the phosphorylation of β‑catenin and N‑cadherin expression. Furthermore, CPT injection significantly suppressed pancreatic tumor growth and peritoneal dissemination of highly metastatic SUIT‑2 pancreatic cancer cells in a mouse orthotopic pancreatic cancer model, without evident toxicity. Moreover, immunohistochemistry analyses demonstrated decreased β‑catenin expression in CPT‑treated pancreatic tumors compared with control tumors. Taken together, these results indicate that CPT reduced the invasiveness and metastasis of bladder cancer cells by suppressing EMT via the mTOR/β‑catenin/N‑cadherin pathway.

Topics: Animals; Antigens, CD; Antineoplastic Agents; beta Catenin; Cadherins; Cell Line, Tumor; Female; Humans; Mice, Inbred BALB C; Mice, Nude; Pancreatic Neoplasms; Phenanthrenes; Pyruvate Dehydrogenase Acetyl-Transferring Kinase; Signal Transduction; Spheroids, Cellular; TOR Serine-Threonine Kinases; Urinary Bladder Neoplasms; Xenograft Model Antitumor Assays

Hydroxycamptothecin (HCPT) is used as a clinical chemotherapy regimen to treat bladder cancer, but more efficacious novel combination treatments are needed.. Cultured bladder cancer cell lines EJ and UMUC3 were treated with triptolide (TPL) and/or HCPT. A flow cytometry approach was used to detect cell cycle phase, apoptosis and reactive oxygen species. Western blotting was used to measure CDK4, CDK6, CyclinD1, catalase, Caspase8 and Bcl-xl protein levels in control, TPL treatment, HCPT treatment and TPL plus HCPT treatment bladder cancer cells. AKT pathway proteins, including AKT and p-AKT, were also detected by western blotting. UMUC3 cells treated with DMSO, HCPT, TPL and HCPT plus TPL were injected subcutaneously into mice (n = 3 per group).. The flow cytometry and western blot results indicated that compared to TPL or HCPT treatment alone, combination treatment of HCPT and TPL significantly induced cell cycle arrest at the G1 phase via suppressing CDK4, CDK6 and CyclinD1 in the EJ and UMUC3 bladder cancer cell lines. HCPT and TPL combination treatment also significantly increased the apoptosis rate and apoptosis-related protein levels (Caspase8 and Bcl-xl). Levels of the AKT pathway-related proteins AKT/p-AKT were significantly lower in EJ and UMUC3 cells treated with TPL and UMUC3 than in those cells treated with TPL or HCPT alone. TPL plus HCPT treatment significantly reduced bladder tumour sizes in vivo on the seventh and tenth days.. Compared to TPL or HCPT treatment alone, TPL plus HCPT combination treatment had significantly better anticancer effects.

Topics: Animals; Antineoplastic Agents; Apoptosis; Camptothecin; Cell Cycle; Cell Line, Tumor; Cell Proliferation; Cell Survival; Diterpenes; Drug Synergism; Epoxy Compounds; Mice, Inbred BALB C; Mice, Nude; Phenanthrenes; Reactive Oxygen Species; Urinary Bladder Neoplasms; Xenograft Model Antitumor Assays

Gemcitabine (GEM) effectively inhibits bladder cancer progression in the clinic, but novel combination treatments using multiple drugs are needed.. The bladder cancer cell lines EJ and UMUC3 were treated with triptolide (TPL) and/or GEM. Tumour cell viability and proliferation were measured using MTT and clonogenic assays, respectively. Flow cytometry and western blotting were used to detect the cell cycle phase, apoptosis, reactive oxygen species (ROS) and the levels of specific relevant proteins. The AKT/GSK3β signalling pathway proteins were also measured by immunofluorescence and western blotting.. The cytotoxicity of the GEM plus TPL combination treatment was stronger than that of GEM or TPL alone. In bladder cancer cell lines, GEM plus TPL induced cell cycle arrest at the G1 phase via suppression of CDK4, CDK6 and cyclins A1 and A2. Significantly increased apoptosis and increases in apoptosis-related proteins (caspase 8 and Bcl-xL) were observed in cells treated with GEM plus TPL. While ROS increased, certain ROS-related proteins (catalase and SOD2) clearly decreased in cells treated with a combination of GEM plus TPL. The AKT/GSK3β signalling pathway was also inhibited more significantly in cells treated with the GEM plus TPL combination than in cells treated with either agent alone.. The combination of GEM plus TPL showed significantly enhanced anticancer effects compared to those of GEM or TPL alone.

Topics: Antineoplastic Combined Chemotherapy Protocols; Apoptosis; Apoptosis Regulatory Proteins; Cell Cycle Proteins; Cell Line, Tumor; Cell Proliferation; Cell Survival; Deoxycytidine; Diterpenes; Dose-Response Relationship, Drug; Drug Synergism; Epoxy Compounds; G1 Phase Cell Cycle Checkpoints; Gemcitabine; Glycogen Synthase Kinase 3 beta; Humans; Phenanthrenes; Proto-Oncogene Proteins c-akt; Reactive Oxygen Species; Signal Transduction; Urinary Bladder Neoplasms

We examined the synergistic antitumor effects of cisplatin combined with triptolide in cisplatin resistant T24R2 bladder cancer cells and investigated possible molecular mechanisms.. T24R2 cells were treated with cisplatin and/or triptolide. Tumor cell proliferation was determined using cell counting and clonogenic assays. The combination index of the synergism between cisplatin and triptolide was calculated. The cell cycle phase and apoptosis rate were determined by flow cytometry. Apoptosis and cell cycle related protein expression were analyzed by Western blot.. The synergistic cytotoxicity effect of cisplatin and triptolide combination treatment was greater than the cytotoxic effect of cisplatin or triptolide alone. Combination treatment also induced cell cycle arrest via cyclin D1 and E1 expression. Apoptosis induced by combination treatment was accompanied by increased expression of caspase-3, 8 and 9, PARP and cytochrome c.. Our results suggest that triptolide synergistically enhanced the antitumor effect of cisplatin in cisplatin resistant human bladder cancer cells. Cisplatin and triptolide combination treatment may be effective for advanced bladder cancer.

Topics: Antineoplastic Agents; Antineoplastic Combined Chemotherapy Protocols; Cisplatin; Diterpenes; Drug Resistance, Neoplasm; Drug Synergism; Epoxy Compounds; Humans; Phenanthrenes; Tumor Cells, Cultured; Urinary Bladder Neoplasms

Triptolide (TPL), a diterpenoid triepoxide purified from the Chinese herb Tripterygium wilfordii Hook F, was tested for its antitumor properties in several model systems. In vitro, TPL inhibited the proliferation and colony formation of tumor cells at extremely low concentrations (2-10 ng/ml) and was more potent than Taxol. Likewise, in vivo, treatment of mice with TPL for 2-3 weeks inhibited the growth of xenografts formed by four different tumor cell lines (B16 melanoma, MDA-435 breast cancer, TSU bladder cancer, and MGC80-3 gastric carcinoma), indicating that TPL has a broad spectrum of activity against tumors that contain both wild-type and mutant forms of p53. In addition, TPL inhibited experimental metastasis of B16F10 cells to the lungs and spleens of mice. The antitumor effect of TPL was comparable or superior with that of conventional antitumor drugs, such as Adriamycin, mitomycin, and cisplatin. Importantly, tumor cells that were resistant to Taxol attributable to the overexpression of the multidrug resistant gene 1 were still sensitive to the effects of TPL. Studies on cultured tumor cells revealed that TPL induced apoptosis and reduced the expression of several molecules that regulate the cell cycle. Taken together, these results suggest that TPL has several attractive features as a new antitumor agent.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Antineoplastic Agents; Breast Neoplasms; Cell Division; Diterpenes; Epoxy Compounds; Female; Humans; Melanoma; Melanoma, Experimental; Mice; Neoplasm Metastasis; Phenanthrenes; Stomach Neoplasms; Tumor Cells, Cultured; Urinary Bladder Neoplasms

Topics: Anti-Obesity Agents; Aristolochia; Aristolochic Acids; Atrophy; Biomarkers; Carcinogens; Carcinoma, Papillary; Carcinoma, Transitional Cell; DNA Adducts; DNA Damage; Drugs, Chinese Herbal; Female; Fibrosis; Humans; Kidney Neoplasms; Kidney Transplantation; Kidney Tubular Necrosis, Acute; Kidney Tubules, Proximal; Male; Neprilysin; Phenanthrenes; Phytotherapy; Plant Preparations; Postoperative Complications; Urinary Bladder Neoplasms

We have shown that the oval cell line OC/CDE 22 can be transformed by the highly carcinogenic fjord-region diol epoxides of benzo[c]phenanthrene. Mutational activation of the ras proto-oncogene family has been proposed to be a critical event in the formation of tumors induced by polycyclic aromatic hydrocarbons. Therefore, we investigated whether in the earlier transformed OC/CDE 22 cells any point mutations were detected in the ras proto-oncogene. The results indicate that the malignant transformation of OC/CDE 22 cells by the 4 stereoisomeric benzo[c]phenanthrene diol epoxides in vitro is independent of activation of the Ha-ras proto-oncogene. In addition, Northern and Western blot analyses revealed no overexpression of the Ha-ras protooncogene in the transformed OC/CDE 22 cell lines. However, transfection of the OC/CDE 22 cells with an activated Ha-ras oncogene malignantly transformed the OC/CDE 22 cells, and the transfected cells served as precursor cells of tumors with a cholangiocellular morphology and phenotype. Our latter finding reinforces the view that OC/CDE 22 cells are committed to the bile duct epithelial cell lineage.

Topics: Animals; Bile Ducts; Carcinogens; Carcinoma; Cell Differentiation; Cell Line; Cell Lineage; Cell Transformation, Neoplastic; Epithelial Cells; Gene Expression Regulation, Neoplastic; Genes, ras; Humans; Liver; Liver Neoplasms, Experimental; Neoplasm Transplantation; Neoplastic Stem Cells; Phenanthrenes; Proto-Oncogene Mas; Rats; Rats, Sprague-Dawley; Transfection; Urinary Bladder Neoplasms

Topics: Acridines; Animals; Benz(a)Anthracenes; Benzopyrenes; Carcinogens; Female; Fluorenes; Mice; Mice, Inbred Strains; Nicotiana; Phenanthrenes; Plants, Toxic; Radioisotopes; Smoke; Smoking; Urinary Bladder Neoplasms