title: Lipoic acid induces p53-independent cell death in colorectal cancer cells and potentiates the cytotoxicity of 5-fluorouracil.
abstract: Alpha-lipoic acid (LA), which plays a pivotal role in mitochondrial energy metabolism, is an endogenous dithiol compound with an array of antioxidative functions. It has been shown that LA triggers cell death in tumor cell lines, whereas non-transformed cells are hardly affected. In the present study, we analyzed the cytotoxicity of LA on colorectal cancer (CRC) cells differing in their p53 status and investigated a putative synergistic effect with the anticancer drug 5-fluorouracil (5-FU). We show that LA induces a dose-dependent decrease in cell viability, which was independent of the p53 status as attested in isogenic p53-proficient and p53-deficient cell lines. This effect was largely attributable to cell death induction as revealed by Annexin-V/PI staining. LA-treated HCT116 cells underwent caspase-dependent and caspase-independent cell death, which was blocked by the pan-caspase inhibitor zVAD and the RIP-kinase inhibitor Necrostatin-1, respectively. In CaCO-2 and HT29 cells, LA induced caspase-dependent cell demise via activation of caspase-9, caspase-3 and caspase-7 with subsequent PARP-1 cleavage as demonstrated by immunoblot analysis, activity assays and pan-caspase inhibition. Interestingly, LA treatment did neither activate p53 nor induced genotoxic effects as shown by lack of DNA strand breaks and phosphorylation of histone 2AX. Finally, we provide evidence that LA increases the cytotoxic effect induced by the anticancer drug 5-FU as revealed by significantly enhanced cell death rates in HCT116 and CaCO-2 cells. Collectively, these findings demonstrate that LA induces CRC cell death independent of their p53 status and potentiates the cytotoxicity of 5-FU without causing DNA damage on its own, which makes it a candidate for tumor therapy.
authors.affiliation: Department of Toxicology, University Medical Center, Obere Zahlbacher Str. 67, 55131, Mainz, Germany.
authors.forename: Bastian
authors.initials: B
authors.lastname: Dörsam
authors.affiliation: Department of Toxicology, University Medical Center, Obere Zahlbacher Str. 67, 55131, Mainz, Germany.
authors.forename: Anja
authors.initials: A
authors.lastname: Göder
authors.affiliation: Department of Toxicology, University Medical Center, Obere Zahlbacher Str. 67, 55131, Mainz, Germany.
authors.forename: Nina
authors.initials: N
authors.lastname: Seiwert
authors.affiliation: Department of Toxicology, University Medical Center, Obere Zahlbacher Str. 67, 55131, Mainz, Germany.
authors.forename: Bernd
authors.initials: B
authors.lastname: Kaina
authors.affiliation: Department of Toxicology, University Medical Center, Obere Zahlbacher Str. 67, 55131, Mainz, Germany. fahrer@uni-mainz.de.
authors.forename: Jörg
authors.initials: J
authors.lastname: Fahrer
mesh_terms: D000964:Antimetabolites, Antineoplastic / Q000008:administration & dosage / Q000494:pharmacology*; D018938:Caco-2 Cells; D020169:Caspases / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D015179:Colorectal Neoplasms / Q000188:drug therapy* / Q000473:pathology; D004305:Dose-Response Relationship, Drug; D004357:Drug Synergism; D005472:Fluorouracil / Q000008:administration & dosage / Q000494:pharmacology*; D045325:HCT116 Cells; D006801:Humans; D008063:Thioctic Acid / Q000494:pharmacology*; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism
chemical_list: D000964:Antimetabolites, Antineoplastic; D016159:Tumor Suppressor Protein p53; D008063:Thioctic Acid; D020169:Caspases; D005472:Fluorouracil
keywords: 5-Fluorouracil; Cell death; Colorectal cancer; Genotoxicity; Lipoic acid; p53