Assessment of pro-apoptotic activity of doxorubicin–transferrin conjugate in cells derived from human solid tumors

• Published in: The international journal of biochemistry & cell biology 2016-01, Vol.70, p.57-67
• Main Authors: Szwed, Marzena, Kania, Katarzyna Dominika, Jozwiak, Zofia
• Format: Article
• Language: English
• Subjects: Anthracyclines; Antineoplastic Agents - pharmacology; Apoptosis - drug effects; Apoptosis - genetics; Apoptosis Inducing Factor - genetics; Apoptosis Inducing Factor - metabolism; bcl-2-Associated X Protein - genetics; bcl-2-Associated X Protein - metabolism; Calpain - genetics; Calpain - metabolism; Caspase 3 - genetics; Caspase 3 - metabolism; Caspase 9 - genetics; Caspase 9 - metabolism; Cell Line, Tumor; Cell Survival - drug effects; DNA Fragmentation - drug effects; Doxorubicin - pharmacology; Drug delivery systems; Drug Resistance, Neoplasm - genetics; Epithelial Cells - drug effects; Epithelial Cells - metabolism; Epithelial Cells - pathology; Gene Expression Regulation, Neoplastic; Hep G2 Cells; Humans; Membrane Potential, Mitochondrial - drug effects; Proto-Oncogene Proteins c-bcl-2 - genetics; Proto-Oncogene Proteins c-bcl-2 - metabolism; Real-Time Polymerase Chain Reaction; Signal Transduction; Solid tumors; Transferrin; Transferrin - pharmacology
• ISSN: 1357-2725; 1878-5875
• DOI: 10.1016/j.biocel.2015.10.020

•DOX–TRF conjugate induces caspase-dependent apoptosis in cancer cells at a low dose.•Human tumor cells are more sensitive to DOX–TRF conjugate application than DOX alone.•DOX–TRF conjugate shows a higher level of pro-apoptotic DNA damage than free DOX. Conjugates of anthracyclines are a new possibility for anticancer agent delivery, which seems to be a very promising alternative to the currently used cancer treatment strategies. In our study, we investigated the ability of a doxorubicin–transferrin (DOX–TRF) conjugate to induce cell death in two solid tumor cell lines: non-small cell lung cancer (A549) and hepatocellular liver carcinoma (HepG2). The observed effects of the DOX–TRF conjugate on these cell cultures were compared with those of free doxorubicin (DOX), a widely used antineoplastic therapeutic agent. Our results provided direct evidence that the investigated conjugate is considerably more cytotoxic to the examined human cancer cell lines than is DOX alone. Moreover, we confirmed that the antitumor efficacy of DOX–TRF conjugate is related to its apoptosis-inducing ability, which was shown during measurements of typical features of programmed cell death. In solid tumor cell lines, the DOX–TRF conjugate induced changes in cellular morphology, mitochondrial membrane potential and caspases-3 and -9 activities. Furthermore, all of the analyzed hallmarks of apoptosis were confirmed by the oligonucleosomal DNA fragmentation assay and by a real-time PCR quantitative study, which displayed the superiority of the conjugate-induced programmed cell death over free drug-triggered cell death.