Development of a therapeutic model of precancerous liver using crocin-coated magnetite nanoparticles

El-Kharrag,Rkia; Amin,Amr; Hisaindee,Soleiman; Greish,Yaser; Karam,Sherif M.

doi:10.3892/ijo.2016.3769

Development of a therapeutic model of precancerous liver using crocin-coated magnetite nanoparticles

Authors:
- Rkia El-Kharrag
- Amr Amin
- Soleiman Hisaindee
- Yaser Greish
- Sherif M. Karam
View Affiliations

Affiliations: Department of Biology, College of Science, UAE University, Al-Ain, United Arab Emirates, Department of Chemistry, College of Science, UAE University, Al-Ain, United Arab Emirates, Department of Anatomy, College of Medicine and Health Sciences, UAE University, Al-Ain, United Arab Emirates
Published online on: November 15, 2016 https://doi.org/10.3892/ijo.2016.3769
Pages: 212-222

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Despite considerable advances in understanding hepatocellular carcinoma, it is one of the common and deadliest cancers worldwide. Hence, increasing efforts are needed for early diagnosis and effective treatments. Saffron has been recently found to inhibit growth of liver cancer in rats. The aim of this study was to develop an effective method for treatment of liver cancer using magnetite nanoparticles (MNPs) coated with crocin, the main active component of saffron. MNPs were prepared and initially coated with dextran and a cross-linker to enhance conjugation of crocin using a modified coprecipitation method. Cultured HepG2 cells and diethylnitrosamine-injected mice were treated with corcin-coated MNPs and analyzed using cell proliferation assay and immunohistochemical analysis, respectively. Treatment of HepG2 cells with crocin-coated MNPs led to a significant inhibition of their growth as compared to control or those treated with free crocin or uncoated MNPs. Histological examinations of the livers of diethylnitrosamine-injected mice revealed several precancerous changes: multiple proliferative hepatic foci, hyper- or dysplastic transformations of bile ducts/ductules, and nuclear atypia associated with polyploidy, karyomegaly, and vacuolation. Immunohistochemistry using antibodies specific for cell proliferation (Ki-67) and apoptosis (M30-CytoDEATH and Bcl-2) revealed their upregulation during development of precancerous lesions. Using antibodies specific for inflammation (cyclooxygenase-2), oxidative stress (glutathione) and angiogenesis (vascular endothelial growth factor) indicated the involvement of multiple signaling pathways in the development of precancerous lesions. Treatment with crocin-coated MNPs was associated with regression of precancerous lesions, significant upregulation of apoptotic cells and downregulation of Bcl-2 labeling and markers of cell proliferation, inflammation, oxidative stress and angiogenesis. In conclusion, crocin-coated MNPs are more effective than free corcin for treatment of liver precancerous lesions in mice. These findings will help to develop new modalities for early detection and treatment of liver precancerous lesions.

View Figures

View References

1	Torre LA, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J and Jemal A: Global cancer statistics, 2012. CA Cancer J Clin. 65:87–108. 2015. View Article : Google Scholar : PubMed/NCBI
2	London W and McGlynn K: Liver cancer. Cancer Epidemiology and Prevention. 3rd. Schottenfeld D and Fraumeni JF: Oxford University Press; New York, NY: pp. 763–786. 2006, View Article : Google Scholar
3	Llovet JM, Ricci S, Mazzaferro V, Hilgard P, Gane E, Blanc JF, de Oliveira AC, Santoro A, Raoul JL, Forner A, et al SHARP Investigators Study Group: Sorafenib in advanced hepatocellular carcinoma. N Engl J Med. 359:378–390. 2008. View Article : Google Scholar : PubMed/NCBI
4	Tietze R, Zaloga J, Unterweger H, Lyer S, Friedrich RP, Janko C, Pöttler M, Dürr S and Alexiou C: Magnetic nanoparticle-based drug delivery for cancer therapy. Biochem Biophys Res Commun. 468:463–470. 2015. View Article : Google Scholar : PubMed/NCBI
5	Block KI, Gyllenhaal C, Lowe L, Amedei A, Amin AR, Amin A, Aquilano K, Arbiser J, Arreola A, Arzumanyan A, et al: Designing a broad-spectrum integrative approach for cancer prevention and treatment. Semin Cancer Biol. 35(Suppl): S276–S304. 2015. View Article : Google Scholar : PubMed/NCBI
6	Kianbakht S and Mozaffari K: Effects of saffron and its active constituents, crocin and safranal on prevention of indomethacin induced gastric ulcers in diabetic and nondiabetic rats. J Med Plants. 8:30–38. 2009.
7	Amin A, Hamza AA, Bajbouj K, Ashraf SS and Daoud S: Saffron: A potential candidate for a novel anticancer drug against hepatocellular carcinoma. Hepatology. 54:857–867. 2011. View Article : Google Scholar : PubMed/NCBI
8	Tamaddonfard E, Farshid AA, Eghdami K, Samadi F and Erfanparast A: Comparison of the effects of crocin, safranal and diclofenac on local inflammation and inflammatory pain responses induced by carrageenan in rats. Pharmacol Rep. 65:1272–1280. 2013. View Article : Google Scholar
9	Khosrojerdi F and Kazemi Noureini S: Study of telomerase activity in cell line MCF7 treated with crocin. Clin Biochem. 44:S1132011. View Article : Google Scholar
10	Escribano J, Alonso GL, Coca-Prados M and Fernández JA: Crocin, safranal and picrocrocin from saffron (Crocus sativus L.) inhibit the growth of human cancer cells in vitro. Cancer Lett. 100:23–30. 1996. View Article : Google Scholar : PubMed/NCBI
11	Singh A and Sahoo SK: Magnetic nanoparticles: A novel platform for cancer theranostics. Drug Discov Today. 19:474–481. 2014. View Article : Google Scholar
12	El-kharrag R, Amin A and Greish YE: Low temperature synthesis of monolithic mesoporous magnetite nanoparticles. Ceram Int. 38:627–634. 2012. View Article : Google Scholar
13	El-kharrag R, Amin A and Greish YE: Synthesis and characterization of mesoporous sodium dodecyl sulfate-coated magnetite nanoparticles. J Ceram Sci Tech. 2:203–210. 2011.
14	Sundar S, Mariappan R and Piraman S: Synthesis and characterization of amine modified magnetite nanoparticles as carriers of curcumin-anticancer drug. Powder Technol. 266:321–328. 2014. View Article : Google Scholar
15	Mosmann T: Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. J Immunol Methods. 65:55–63. 1983. View Article : Google Scholar : PubMed/NCBI
16	Mejías R, Pérez-Yagüe S, Gutiérrez L, Cabrera LI, Spada R, Acedo P, Serna CJ, Lázaro FJ, Villanueva A, Morales MP, et al: Dimercaptosuccinic acid-coated magnetite nanoparticles for magnetically guided in vivo delivery of interferon gamma for cancer immunotherapy. Biomaterials. 32:2938–2952. 2011. View Article : Google Scholar : PubMed/NCBI
17	Dar RA, Brahaman PK, Tiwari S and Pitre KS: Indirect electrochemical analysis of crocin in phytochemical sample. E-J Chem. 9:918–925. 2012. View Article : Google Scholar
18	Kumar SR, Priyatharshni S, Babu VN, Mangalaraj D, Viswanathan C, Kannan S and Ponpandian N: Quercetin conjugated superparamagnetic magnetite nanoparticles for in-vitro analysis of breast cancer cell lines for chemotherapy applications. J Colloid Interface Sci. 436:234–242. 2014. View Article : Google Scholar : PubMed/NCBI
19	Dawidczyk CM, Kim C, Park JH, Russell LM, Lee KH, Pomper MG and Searson PC: State-of-the-art in design rules for drug delivery platforms: Lessons learned from FDA-approved nanomedicines. J Control Release. 187:133–144. 2014. View Article : Google Scholar : PubMed/NCBI
20	Molday RS and MacKenzie D: Immunospecific ferromagnetic iron-dextran reagents for the labeling and magnetic separation of cells. J Immunol Methods. 52:353–367. 1982. View Article : Google Scholar : PubMed/NCBI
21	Amin A, Bajbouj K, Koch A, Gandesiri M and Schneider-Stock R: Defective autophagosome formation in p53-null colorectal cancer reinforces crocin-induced apoptosis. Int J Mol Sci. 16:1544–1561. 2015. View Article : Google Scholar : PubMed/NCBI
22	Aung HH, Wang CZ, Ni M, Fishbein A, Mehendale SR, Xie JT, Shoyama CY and Yuan CS: Crocin from Crocus sativus possesses significant anti-proliferation effects on human colorectal cancer cells. Exp Oncol. 29:175–180. 2007.PubMed/NCBI
23	Veiseh O, Gunn JW and Zhang M: Design and fabrication of magnetic nanoparticles for targeted drug delivery and imaging. Adv Drug Deliv Rev. 62:284–304. 2010. View Article : Google Scholar :
24	Rumsby PC, Barrass NC, Phillimore HE and Evans JG: Analysis of the Ha-ras oncogene in C3H/He mouse liver tumours derived spontaneously or induced with diethylnitrosamine or phenobarbitone. Carcinogenesis. 12:2331–2336. 1991. View Article : Google Scholar : PubMed/NCBI
25	He XY, Smith GJ, Enno A and Nicholson RC: Short-term diethylnitrosamine-induced oval cell responses in three strains of mice. Pathology. 26:154–160. 1994. View Article : Google Scholar : PubMed/NCBI
26	Nakatani T, Roy G, Fujimoto N, Asahara T and Ito A: Sex hormone dependency of diethylnitrosamine-induced liver tumors in mice and chemoprevention by leuprorelin. Jpn J Cancer Res. 92:249–256. 2001. View Article : Google Scholar : PubMed/NCBI
27	Lowe SW and Lin AW: Apoptosis in cancer. Carcinogenesis. 21:485–495. 2000. View Article : Google Scholar : PubMed/NCBI
28	Lee GH: Correlation between Bcl-2 expression and histopathology in diethylnitrosamine-induced mouse hepatocellular tumors. Am J Pathol. 151:957–961. 1997.PubMed/NCBI
29	Satoh K, Kitahara A, Soma Y, Inaba Y, Hatayama I and Sato K: Purification, induction, and distribution of placental glutathione transferase: A new marker enzyme for preneoplastic cells in the rat chemical hepatocarcinogenesis. Proc Natl Acad Sci USA. 82:3964–3968. 1985. View Article : Google Scholar : PubMed/NCBI
30	Gallo O, Franchi A, Magnelli L, Sardi I, Vannacci A, Boddi V, Chiarugi V and Masini E: Cyclooxygenase-2 pathway correlates with VEGF expression in head and neck cancer. Implications for tumor angiogenesis and metastasis. Neoplasia. 3:53–61. 2001. View Article : Google Scholar : PubMed/NCBI
31	Tang TC, Poon RT, Lau CP, Xie D and Fan ST: Tumor cyclooxygenase-2 levels correlate with tumor invasiveness in human hepatocellular carcinoma. World J Gastroenterol. 11:1896–1902. 2005. View Article : Google Scholar : PubMed/NCBI
32	Sivaramakrishnan V and Niranjali Devaraj S: Morin regulates the expression of NF-kappaB-p65, COX-2 and matrix metalloproteinases in diethylnitrosamine induced rat hepatocellular carcinoma. Chem Biol Interact. 180:353–359. 2009. View Article : Google Scholar : PubMed/NCBI
33	Kawabata K, Tung NH, Shoyama Y, Sugie S, Mori T and Tanaka T: Dietary crocin inhibits colitis and colitis-associated colorectal carcinogenesis in male ICR mice. Evid Based Complement Alternat Med. 2012:8204152012. View Article : Google Scholar
34	Lee SH, Jeong D, Han YS and Baek MJ: Pivotal role of vascular endothelial growth factor pathway in tumor angiogenesis. Ann Surg Treat Res. 89:1–8. 2015. View Article : Google Scholar : PubMed/NCBI
35	Maggi L, Carmona M, Zalacain A, Tomé MM, Murcia MA and Alonso GL: Parabens as agents for improving crocetin esters' shelf-life in aqueous saffron extracts. Molecules. 14:1160–1170. 2009. View Article : Google Scholar : PubMed/NCBI