4-methylene-2-octyl-5-oxofuran-3-carboxylic-acid and Prostatic-Neoplasms

Many common human cancers, including colon, prostate and breast cancer, express high levels of fatty acid synthase compared to normal human tissues. This elevated expression is associated with protection against apoptosis, increased metastasis and poor prognosis. Inhibitors of fatty acid synthase, such as the cerulenin synthetic analog C75, decrease prostate cancer cell proliferation, increase apoptosis and decrease tumor growth in experimental models. Although radiotherapy is widely used in the treatment of prostate cancer patients, the risk of damage to neighboring normal organs limits the radiation dose that can be delivered. In this study, we examined the potential of fatty acid synthase inhibition to sensitize prostate cancer cells to radiotherapy. The efficacy of C75 alone or in combination with X irradiation was examined in monolayers and in multicellular tumor spheroids. Treatment with C75 alone decreased clonogenic survival, an effect that was abrogated by the antioxidant. C75 treatment also delayed spheroid growth in a concentration-dependent manner. The radiosensitizing effect of C75 was indicated by combination index values between 0.65 and 0.71 and the reduced surviving fraction of clonogens, in response to 2 Gy X irradiation, from 0.51 to 0.30 and 0.11 in the presence of 25 and 35 μM C75, respectively. This increased sensitivity to radiation was reduced by the presence of the antioxidant. The C75 treatment also enhanced the spheroid growth delay induced by X irradiation in a supra-additive manner. The level of radiation-induced apoptosis in prostate cancer cells was increased further by C75, which induced cell cycle arrest in the G2/M phase, but only at a concentration greater than that required for radiosensitization. Radiation-induced G2/M blockade was not affected by C75 treatment. These results suggest the potential use of fatty acid synthase inhibition to enhance the efficacy of radiotherapy of prostate carcinoma and that C75-dependent cell cycle arrest is not responsible for its radiosensitizing effect.

Topics: 4-Butyrolactone; Antioxidants; Apoptosis; Cell Cycle; Cell Line, Tumor; Cell Movement; Dose-Response Relationship, Drug; Enzyme Inhibitors; Fatty Acid Synthases; Humans; Male; Prostatic Neoplasms; Radiation Tolerance; Spheroids, Cellular; X-Rays

Inhibition of FASN has emerged as a promising therapeutic target in cancer, and numerous inhibitors have been investigated. However, severe pharmacological limitations have challenged their clinical testing. The synthetic FASN inhibitor triclosan, which was initially developed as a topical antibacterial agent, is merely affected by these pharmacological limitations. Yet, little is known about its mechanism in inhibiting the growth of cancer cells. Here we compared the cellular and molecular effects of triclosan in a panel of eight malignant and non-malignant prostate cell lines to the well-known FASN inhibitors C75 and orlistat, which target different partial catalytic activities of FASN. Triclosan displayed a superior cytotoxic profile with a several-fold lower IC50 than C75 or orlistat. Structure-function analysis revealed that alcohol functionality of the parent phenol is critical for inhibitory action. Rescue experiments confirmed that end product starvation was a major cause of cytotoxicity. Importantly, triclosan, C75 and orlistat induced distinct changes to morphology, cell cycle, lipid content and the expression of key enzymes of lipid metabolism, demonstrating that inhibition of different partial catalytic activities of FASN activates different metabolic pathways. These finding combined with its well-documented pharmacological safety profile make triclosan a promising drug candidate for the treatment of prostate cancer.

Topics: 3T3 Cells; 4-Butyrolactone; Animals; Anti-Infective Agents, Local; Antineoplastic Agents; Apoptosis; Cell Line, Tumor; Drug Repositioning; Fatty Acid Synthase, Type I; Fatty Acid Synthesis Inhibitors; G1 Phase Cell Cycle Checkpoints; Humans; Lactones; Lipid Metabolism; Male; Mice; Orlistat; Prostatic Neoplasms; Structure-Activity Relationship; Triclosan

Positron emission tomography (PET) imaging with copper (II)-diacetyl-bis(N4-Methylthiosemicarbazone)(Cu-ATSM) for delineating hypoxia has provided valuable clinical information, but investigations in animal models of prostate cancer have shown some inconsistencies. As a defense mechanism in prostate cancer cells, the fatty acid synthesis pathway harnesses its oxidizing power for improving the redox balance despite conditions of extreme hypoxia, potentially altering Cu-ATSM hypoxia selectivity.. Human prostate tumor-cultured cell lines (PC-3, 22Rv1, LNCaP and LAPC-4), were treated with a fatty acid synthase (FAS) inhibitor (C75, 100 microM) under anoxia. The 64Cu-ATSM uptake in these treated cells and nontreated anoxic cells was then examined. Fatty acid synthase expression level in each cell line was subsequently quantified by ELISA. An additional study was performed in PC-3 cells to examine the relationship between the restoration of 64Cu-ATSM hypoxia selectivity and the concentration of C75 (100, 20, 4 or 0.8 microM) administered to the cells.. Inhibition of fatty acid synthesis with C75 resulted in a significant increase in 64Cu-ATSM retention in prostate tumor cells in vitro under anoxia over 60 min. Inhibition studies demonstrated higher uptake values of 20.9+/-3.27%, 103.0+/-32.6%, 144.2+/-32.3% and 200.1+/-79.3% at 15 min over control values for LAPC-4, PC-3, LNCaP and 22Rv1 cells, respectively. A correlation was seen (R2=.911) with FAS expression plotted against percentage change in 64Cu-ATSM uptake with C75 treatment.. Although Cu-ATSM has clinical relevance in the PET imaging of hypoxia in many tumor types, its translation to the imaging of prostate cancer may be limited by the overexpression of FAS associated with prostatic malignancies.

Topics: 4-Butyrolactone; Cell Hypoxia; Cell Line, Tumor; Coordination Complexes; Copper Radioisotopes; Dose-Response Relationship, Drug; Drug Synergism; Fatty Acid Synthases; Gene Expression; Humans; Male; Organometallic Compounds; Positron-Emission Tomography; Prostatic Neoplasms; Radiopharmaceuticals; Thiosemicarbazones

Fatty acid synthase (FAS) performs the anabolic conversion of dietary carbohydrate or protein to fat. FAS expression is low in most normal tissues, but is elevated in many human cancers, including androgen-sensitive and androgen-independent prostate cancer.. Immunohistochemical evaluation of FAS expression was performed in human prostate cancer specimens under various states of androgen ablation. In vitro and in vivo prostate cancer models were evaluated for FAS expression and activity under androgenic and androgen-depleted conditions, and were tested for sensitivity to antimetabolite drugs that target fatty acid synthesis.. While FAS expression in the prostate was androgen responsive, it persisted or was reactivated in human prostate carcinoma after androgen ablation, and was high in 82% of lethal tumors examined at autopsy. Similar patterns of FAS expression and fatty acid synthesis were seen in cell culture and xenograft models of human prostate cancer. Pharmacologic inhibition of FAS resulted in a dose-dependent reduction of tumor growth in these models, including fourfold inhibition of an androgen-independent human prostate cancer xenograft with little associated toxicity.. The data suggest that FAS expression/FA synthesis provides an important functional aspect of the malignant phenotype in prostate cancer, perhaps supporting cell growth or survival. FAS expression may be upregulated by alternate signaling pathways important for prostate cancer growth under androgen withdrawal. The re-emergence of FAS expression and activity during the development of androgen independence demonstrate that FAS may serve as a novel target for antimetabolite therapy in prostate cancer.

Topics: 4-Butyrolactone; Androgens; Animals; Antibodies, Monoclonal; Antineoplastic Agents; Blotting, Western; Chromatography, Thin Layer; Enzyme Inhibitors; Fatty Acid Synthases; Humans; Immunohistochemistry; Lipids; Male; Mice; Mice, Nude; Neoplasms, Hormone-Dependent; Prostatic Neoplasms; Random Allocation; Tumor Cells, Cultured; Xenograft Model Antitumor Assays