In vitro and in vivo suppression of hepatocellular carcinoma growth by chitosan nanoparticles.
Chitosan nanoparticles (CNP), a kind of widely used drug carrier, have shown potent cytotoxic effects on various tumour cell lines in vitro and in vivo. This study sought to evaluate the antitumour effect of CNP on growth of human hepatocellular carcinoma (BEL7402) and the possible mechanisms involved. Cells were grown in the absence and presence of various concentrations of CNP with mean particle size of about 40nm. Cell viability, ultrastructural changes, surface charge, mitochondrial membrane potential, reactive oxygen species (ROS) generation, lipid peroxidation, DNA fragmentation and fatty acid composition were analysed by MTT assay, electron microscopy, zetasizer analysis, flow cytometry, spectrophotometric thiobarbituric (TBA) assays, DNA agarose gel electrophoresis and GC/MS respectively. For in vivo experiments, male BABL/c nude mice were implanted with BEL7402 cells subcutaneously to establish human hepatoma model. Chitosan, saline, and CNP with different mean particle size (40, 70 and 100nm) were administrated by oral administration (1mg/kg body weight). Tumour and body weight were measured, morphologic changes of tumour and liver tissues were studied under electron microscope. In vitro, CNP exhibited high antitumour activities with an IC(50) value of 15.01microg/ml, 6.19microg/ml and 0.94microg/ml after treatment for 24h, 48h and 72h respectively. CNP could induce cell necrosis observed by electron microscope and DNA fragmentation. The antitumour mechanism was mediated by neutralisation of cell surface charge, decrease of mitochondrial membrane potential and induction of lipid peroxidation. The tumour growth inhibitory rates on BEL7402 cells in nude mice treated with chitosan and CNP with different mean particle size (40, 70 and 100nm) were 24.07%, 61.69%, 58.98% and 34.91% respectively. Typical necrotic morphological changes of tumour tissues and no liver abnormalities were found under electron microscope. In this paper, results show a strong antitumour effect of CNP on human hepatoma cell line BEL7402 in vitro and in vivo. These findings suggest that CNP could be a kind of promising agent for further evaluations in the treatment of hepatocellular carcinoma.
Qi
Lifeng
L
Zhejiang University, Nano-biology Lab of Animal Science College, Hangzhou 310029, PR China. lifengqi01@hotmail.com
Xu
Zirong
Z
Chen
Minli
M
D000818:Animals; D006528:Carcinoma, Hepatocellular / Q000000981:diagnostic imaging / Q000473:pathology / Q000628:therapy*; D002470:Cell Survival; D048271:Chitosan / Q000627:therapeutic use*; D053836:DNA Degradation, Necrotic; D053938:DNA Fragmentation; D005227:Fatty Acids / Q000378:metabolism; D006801:Humans; D015227:Lipid Peroxidation; D008113:Liver Neoplasms / Q000000981:diagnostic imaging / Q000473:pathology / Q000628:therapy*; D008297:Male; D053078:Membrane Potential, Mitochondrial; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D008854:Microscopy, Electron; D053758:Nanoparticles / Q000627:therapeutic use*; D011897:Random Allocation; D017382:Reactive Oxygen Species / Q000378:metabolism; D014407:Tumor Cells, Cultured; D014463:Ultrasonography
D005227:Fatty Acids; D017382:Reactive Oxygen Species; D048271:Chitosan