in vitro and in vivo suppression of he pa to cellular

In vitro and in vivo suppression of hepatocellular carcinoma growth by chitosan nanoparticles Lifeng Qi a,c, *, Zirong Xu a , Minli Chen b a Zhejiang University, Nano-biology Lab of Animal Science College, Hangzhou 310029, PR China b Centre of Lab Animal Research, Zhejiang College of Traditional Chinese Medicine, Hangzhou 310053, PR China c Orthopaedic Research Lab, Department of Orthopaedics, School of Medicine, West Virginia University, Morgantown 26506-9196, USA A R T I C L E I N F O Article history: Receiv ed 3 April 2006 Received in revised form 10 August 2006 Accepte d 31 August 2006 Available online 17 October 2006 Keywords: Hepatocellular carcinoma Chitosan nanoparticles Antitumour A B S T R A C T Chitosan nanoparticles (CNP), a kind of widely used drug carrier, have shown potent cyto- toxic 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 pos sible mech ani sms inv olv ed. Cel ls were gro wn in the abs enceand pre sence of vari ous concentrations of CNP with mean particle size of about 40 nm. Cell viability, ultrastructural changes, surface charge, mitochondrial membrane potential, reactive oxygen species (ROS) generation, lipid peroxidation, DNA fragmentation and fatty acid composition were ana- lysed by MTT assay, electron microscopy , zetasizer analysis, flow cytometry, spectrophoto- metr ic thiobarbituric (TBA) ass ays, DNA agarose gel el ect rophor esis 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 100 nm) were administrated by oral administration (1 mg/kg body weight). Tumour and body weight were measured, morphologic changes of tumou r and livertissues were studie d underelectron micros cope. In vit ro , CNPexhib ited hig h ant itu mou r acti vit ies wit h an IC 50 val ue of 15. 01 lg/ml, 6.19 lg/ ml and 0.94 lg/ml aft er tre at- ment for24 h, 48 h and72 h resp ecti vel y . CNP cou ld induce cell nec ros is obs erved byelectr on microscope and DNA fragmentation. The antitumo ur mechanism was mediat ed by neutral- isation of cell surface charge, decrease of mitochondrial membrane potential and induction of lipid per oxi dati on.The tumourgrowth inhibitor y rates on BEL7 402 cells in nude mice trea- tedwith chi tosan and CNPwith dif fere nt mea n part icl e siz e (40,70 and 100 nm)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 invitro and in vivo. These finding s sug gestthat CNPcould be a kin d of promis ingagent forfur- ther evaluations in the treatment of hepatocellular carcinoma. Ó 2006 Elsevier Ltd. All rights reserved. 1. In trod uction Hepatocellular carcinoma (HCC) is one of the most common malignancies worldwide. 1 Chemotherapy is one of the most important treatments currently available for cancer diseases. T reatment of patients with HCC remai ns a clinica l chall enge due to the disappointing effects of most chemotherapies. 2 The efficacy of chemotherapy is limited and patients have 0959-80 49/$ - see front matter Ó 2006 Elsevier Ltd. All rights reserved. doi:10.1016/j.ejca.2006.08.029 * Corresponding author: Tel.: +1 304 293 1065; fax: +1 304 293 7070. E-mail address: [email protected] (L. Qi). E U R O P E A N J O U R N A L O F C A N C E R 43 (2007) 184 – 1 9 3 available at www.sciencedirect.com journal homepage: www.ejconline.com

in vitro and in vivo suppression of he pa to cellular

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