Biodegradable polymer nanoparticle drug delivery systems provide targeted drug delivery, improved pharmacokinetic and biodistribution, enhanced drug stability and fewer side effects. These drug delivery systems are widely used for delivering cytotoxic agents. In the present study, we synthesized GC/5-FU nanoparticles by combining galactosylated chitosan (GC) material with 5-FU, and tested its effect on liver cancer in vitro and in vivo. The in vitro anti-cancer effects of this sustained release system were both dose- and time-dependent, and demonstrated higher cytotoxicity against hepatic cancer cells than against other cell types. The distribution of GC/5-FU in vivo revealed the greatest accumulation in hepatic cancer tissues. GC/5-FU significantly inhibited tumor growth in an orthotropic liver cancer mouse model, resulting in a significant reduction in tumor weight and increased survival time in comparison to 5-FU alone. Flow cytometry and TUNEL assays in hepatic cancer cells showed that GC/5-FU was associated with higher rates of G0–G1 arrest and apoptosis than 5-FU. Analysis of apoptosis pathways indicated that GC/5-FU upregulates p53 expression at both protein and mRNA levels. This in turn lowers Bcl-2/Bax expression resulting in mitochondrial release of cytochrome C into the cytosol with subsequent caspase-3 activation. Upregulation of caspase-3 expression decreased poly ADP-ribose polymerase 1 (PARP-1) at mRNA and protein levels, further promoting apoptosis. These findings indicate that sustained release of GC/5-FU nanoparticles are more effective at targeting hepatic cancer cells than 5-FU monotherapy in the mouse orthotropic liver cancer mouse model.
References
[1]
Wilson B, Ambika TV, Dharmesh KPR, Jenita JL, Priyadarshini SR (2012) Nanoparticles based on albumin: Preparation, characterization and the use for 5-flurouracil delivery. Int J Biol Macromol. Jul 2012 epub ahead of print.
[2]
Ferlay J, Shin HR, Bray F, Forman D, Mathers C, et al. (2010) Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008. Int J Cancer 127: 2893–2917.
[3]
Jemal A, Bray F (2011) Center MM, Ferlay J, Ward E, et al (2011) Global cancer statistics. CA Cancer J Clin 61: 69–90.
[4]
Carr BI (2004) Hepatocellular carcinoma: current management and future trends. Gastroenterology 127: S218–S224.
[5]
Hirai T, Korogi Y, Ono K, Maruoka K, Harada K, et al. (2001) Intraarterial chemotherapy or chemoembolization for locally advanced and/or recurrent hepatic tumors: evaluation of the feeding artery with an interventional CT system. Cardiovasc Intervent Radiol 24: 176–179.
[6]
Burns ER, Beland SS (1983) Induction by 5-fluorouracil of a major phase difference in the circadian profiles of DNA synthesis between the Ehrlich ascites carcinoma and five normal organs. Cancer Lett 20: 235–239.
[7]
Kodama Y, Fumoto S, Nishi J, Nakashima M, Sasaki H, et al. (2008) Absorption and distribution characteristics of 5-fluorouracil (5-FU) after an application to the liver surface in rats in order to reduce systemic side effects. Biol Pharm Bull 31: 1049–1052.
[8]
Wettergren Y, Carlsson G, Odin E, Gustavsson B (2012) Pretherapeutic uracil and dihydrouracil levels of colorectal cancer patients are associated with sex and toxic side effects during adjuvant 5-fluorouracil-based chemotherapy. Cancer 118: 2935–2943.
[9]
Bansal SS, Goel M, Aqil F, Vadhanam MV, Gupta RC (2011) Advanced drug delivery systems of curcumin for cancer chemoprevention. Cancer Prev Res (Phila) 4: 1158–1171.
[10]
Zhang W, Zhang Z, Zhang Y (2011) The application of carbon nanotubes in target drug delivery systems for cancer therapies. Nanoscale Res Lett 6: 555.
[11]
Saha RN, Vasanthakumar S, Bende G, Snehalatha M (2010) Nanoparticulate drug delivery systems for cancer chemotherapy. Mol Membr Biol 27: 215–231.
[12]
De Souza R, Zahedi P, Allen CJ, Piquette-Miller M (2010) Polymeric drug delivery systems for localized cancer chemotherapy. Drug Deliv 17: 365–375.
[13]
Alvarez-Lorenzo C, Sosnik A, Concheiro A (2011) PEO-PPO block copolymers for passive micellar targeting and overcoming multidrug resistance in cancer therapy. Curr Drug Targets 12: 1112–1130.
[14]
Danhier F, Feron O, Preat V (2010) To exploit the tumor microenvironment: Passive and active tumor targeting of nanocarriers for anti-cancer drug delivery. J Control Release 148: 135–146.
[15]
Singh G, Ghosh B, Kaushalkumar D, Somsekhar V (2008) Screening of venlafaxine hydrochloride for transdermal delivery: passive diffusion and iontophoresis. AAPS Pharm Sci Tech 9: 791–797.
[16]
Moghimi SM, Hunter AC, Murray JC (2001) Long-circulating and target-specific nanoparticles: theory to practice. Pharmacol Rev 53: 283–318.
[17]
Lee JB, Zhang K, Tam YY, Tam YK, Belliveau NM, et al. (2011) Lipid nanoparticle siRNA systems for silencing the androgen receptor in human prostate cancer in vivo. Int J Cancer 131: E781–90.
[18]
Ernsting MJ, Tang WL, MacCallum NW, Li SD (2012) Preclinical pharmacokinetic, biodistribution, and anti-cancer efficacy studies of a docetaxel-carboxymethylcellulose nanoparticle in mouse models. Biomaterials 33: 1445–1454.
[19]
Burke AR, Singh RN, Carroll DL, Wood JC, D’Agostino RJ, et al. (2012) The resistance of breast cancer stem cells to conventional hyperthermia and their sensitivity to nanoparticle-mediated photothermal therapy. Biomaterials 33: 2961–2970.
[20]
Arya G, Vandana M, Acharya S, Sahoo SK (2011) Enhanced antiproliferative activity of Herceptin (HER2)-conjugated gemcitabine-loaded chitosan nanoparticle in pancreatic cancer therapy. Nanomedicine 7: 859–870.
[21]
Sanpui P, Chattopadhyay A, Ghosh SS (2011) Induction of apoptosis in cancer cells at low silver nanoparticle concentrations using chitosan nanocarrier. ACS Appl Mater Interfaces 3: 218–228.
[22]
Guan M, Zhou Y, Zhu QL, Liu Y, Bei YY, et al.. (2012) N-trimethyl chitosan nanoparticle-encapsulated lactosyl-norcantharidin for liver cancer therapy with high targeting efficacy. Nanomedicine. 2012 Feb epub ahead of print.
[23]
Wadher KJ, Kakde RB, Umekar MJ (2011) Formulation and evaluation of a sustained-release tablets of metformin hydrochloride using hydrophilic synthetic and hydrophobic natural polymers. Indian J Pharm Sci 73: 208–215.
[24]
Tataru G, Popa M, Costin D, Desbrieres J (2012) Microparticles based on natural and synthetic polymers for ophthalmic applications. J Biomed Mater Res A 100: 1209–1220.
[25]
Kim TH, Park IK, Nah JW, Choi YJ, Cho CS (2004) Galactosylated chitosan/DNA nanoparticles prepared using water-soluble chitosan as a gene carrier. Biomaterials 25: 3783–3792.
[26]
Fallon RJ, Danaher M, Saxena A (1994) The asialoglycoprotein receptor is associated with a tyrosine kinase in HepG2 cells. J Biol Chem 269: 26626–26629.
[27]
Cheng M, Li Q, Wan T, Hong X, Chen H, et al. (2011) Synthesis and efficient hepatocyte targeting of galactosylated chitosan as a gene carrier in vitro and in vivo. J Biomed Mater Res B Appl Biomater 99: 70–80.
[28]
Ruppova K, Wsolova L, Urbancikova M, Slamenova D (2000) Comparison of three in vitro assays at evaluation of IC50 of acetylsalicylic acid, ferrous sulfate, amitriptyline, methanol, isopropanol and ethylene glycol in human cancer cells He La. Neoplasma 47: 172–176.
[29]
Gould WR, McClanahan TB, Welch KM, Baxi SM, Saiya-Cork K, et al. (2006) Inhibitors of blood coagulation factors Xa and IIa synergize to reduce thrombus weight and thrombin generation in vivo and in vitro. J Thromb Haemost 4: 834–841.
[30]
Chang CJ, Chen YH, Huang KW, Cheng HW, Chan SF, et al. (2007) Combined GM-CSF and IL-12 gene therapy synergistically suppresses the growth of orthotopic liver tumors. Hepatology 45: 746–754.
[31]
Dung TD, Chang HC, Chen CY, Peng WH, Tsai CH, et al. (2011) Zanthoxylum avicennae extracts induce cell apoptosis through protein phosphatase 2A activation in HA22T human hepatocellular carcinoma cells and block tumor growth in xenografted nude mice. Int J Mol Med 28: 927–936.
[32]
Khurana A, McKean H, Kim H, Kim SH, Maguire J, et al. (2012) Silencing of HSulf-2 expression in MCF10DICS.com cells attenuate ductal carcinoma in situ progression to invasive ductal carcinoma in vivo. Breast Cancer Res 14: R43.
[33]
Rojiani MV, Alidina J, Esposito N, Rojiani AM (2010) Expression of MMP-2 correlates with increased angiogenesis in CNS metastasis of lung carcinoma. Int J Clin Exp Pathol 3: 775–781.
[34]
Shi Y, Zheng L, Luo G, Wei J, Zhang J, et al. (2011) Expression of zinc finger 23 gene in human hepatocellular carcinoma. Anticancer Res 31: 3595–3599.
[35]
Li P, Wang SS, Liu H, Li N, McNutt MA, et al. (2011) Elevated serum alpha fetoprotein levels promote pathological progression of hepatocellular carcinoma. World J Gastroenterol 17: 4563–4571.
[36]
Li DC, Zhong XK, Zeng ZP, Jiang JG, Li L, et al. (2009) Application of targeted drug delivery system in Chinese medicine. J Control Release 138: 103–112.
[37]
Whitesides GM (2003) The ‘right’ size in nanobiotechnology. Nat Biotechnol 21: 1161–1165.
[38]
Marcucci F, Lefoulon F (2004) Active targeting with particulate drug carriers in tumor therapy: fundamentals and recent progress. Drug Discov Today 9: 219–228.
[39]
Alex SM, Rekha MR, Sharma CP (2011) Spermine grafted galactosylated chitosan for improved nanoparticle mediated gene delivery. Int J Pharm 410: 125–137.
[40]
Jiang H, Wu H, Xu YL, Wang JZ, Zeng Y (2011) Preparation of galactosylated chitosan/tripolyphosphate nanoparticles and application as a gene carrier for targeting SMMC7721 cells. J Biosci Bioeng 111: 719–724.
[41]
Peer D, Karp JM, Hong S, Farokhzad OC, Margalit R, et al. (2007) Nanocarriers as an emerging platform for cancer therapy. Nat Nanotechnol 2: 751–760.
[42]
Spiess M (1990) The asialoglycoprotein receptor: a model for endocytic transport receptors. Biochemistry 29: 10009–10018.
[43]
Weigel PH, Yik JH (2002) Glycans as endocytosis signals: the cases of the asialoglycoprotein and hyaluronan/chondroitin sulfate receptors. Biochim Biophys Acta 1572: 341–363.
[44]
Ashwell G, Harford J (1982) Carbohydrate-specific receptors of the liver. Annu Rev Biochem 51: 531–554.
[45]
Kim JH, Kim YS, Park K, Lee S, Nam HY, et al. (2008) Antitumor efficacy of cisplatin-loaded glycol chitosan nanoparticles in tumor-bearing mice. J Control Release 127: 41–49.
[46]
Managit C, Kawakami S, Yamashita F, Hashida M (2005) Uptake characteristics of galactosylated emulsion by HepG2 hepatoma cells. Int J Pharm 301: 255–261.
[47]
Wang SL, Yu FB, Jiang TY, Sun CS, Wang T, et al. (2008) Design and synthesis of novel galactosylated polymers for liposomes as gene drug carriers targeting the hepatic asialoglycoprotein receptor. J Drug Target 16: 233–242.
[48]
Gao S, Chen J, Dong L, Ding Z, Yang YH, et al. (2005) Targeting delivery of oligonucleotide and plasmid DNA to hepatocyte via galactosylated chitosan vector. Eur J Pharm Biopharm 60: 327–334.
[49]
Sasaki K, Tsuno NH, Sunami E, Tsurita G, Kawai K, et al. (2010) Chloroquine potentiates the anti-cancer effect of 5-fluorouracil on colon cancer cells. BMC Cancer 10: 370.
[50]
Huang Z, Guo KJ, Guo RX, He SG (2007) Effects of 5-fluouracil combined with sulfasalazine on human pancreatic carcinoma cell line BxPC-3 proliferation and apoptosis in vitro. Hepatobiliary Pancreat Dis Int 6: 312–320.
[51]
Parikh N, Koshy C, Dhayabaran V, Perumalsamy LR, Sowdhamini R, et al. (2007) The N-terminus and alpha-5, alpha-6 helices of the pro-apoptotic protein Bax, modulate functional interactions with the anti-apoptotic protein Bcl-xL. BMC Cell Biol 8: 16.
[52]
Avraam K, Pavlakis K, Papadimitriou C, Vrekoussis T, Panoskaltsis T, et al. (2011) The prognostic and predictive value of ERCC-1, p53, bcl-2 and bax in epithelial ovarian cancer. Eur J Gynaecol Oncol 32: 516–520.
[53]
Lee JS, Jung WK, Jeong MH, Yoon TR, Kim HK (2012) Sanguinarine induces apoptosis of HT-29 human colon cancer cells via the regulation of Bax/Bcl-2 ratio and caspase-9-dependent pathway. Int J Toxicol 31: 70–77.
[54]
Choi BH, Kim W, Wang QC, Kim DC, Tan SN, et al. (2008) Kinetin riboside preferentially induces apoptosis by modulating Bcl-2 family proteins and caspase-3 in cancer cells. Cancer Lett 261: 37–45.
