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2017 | 64 | 1 | 25-33
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Biochemical and molecular evidences for the antitumor potential of Ginkgo biloba leaves extract in rodents

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Hepatocellular carcinoma (HCC) is one of the deadliest primary cancers, with a 5-year survival rate of 10% or less. This study was undertaken to elucidate the underlying biochemical and molecular mechanisms in favor of N-nitrosodiethylamine-induced hepatocellular carcinoma. Furthermore, the aim of this work was extended to explore the efficacy of Ginkgo biloba leaves extract in deterioration of HCC in rats. In the current study, HCC group experienced significant downregulation of ING-3 gene expression and upregulation of Foxp-1 gene expression in liver. Treatment of HCC groups with Ginkgo biloba leaves extract resulted in upregulation of ING-3 and downregulation of Foxp-1 gene expression in liver. In addition, there was significant increase in serum alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA) and glypican-3 (GPC-3) levels in HCC group versus the negative control group. In contrast, the groups with HCC subjected to either high or low dose of Ginkgo biloba leaves extract elicited significant reduction (P<0.05) of AFP, CEA and GPC-3 in serum compared to the untreated HCC rats. Besides, histological examination of liver tissue sections of rats in HCC group revealed typical anaplasia. Interestingly, treatment with Ginkgo biloba leaves extract elicited marked improvement in the histological feature of liver tissue in HCC groups. In conclusion, this research indicated that the carcinogenic potency of N-nitrosodiethylamine targeted multiple systems on the cellular and molecular levels. In addition, the results of the current study shed light on the promising anticancer activity of Ginkgo biloba leaves extract in treatment of hepatocellular carcinoma induced chemically in the experimental model through its apoptotic and antiproliferative properties.
Physical description
  • Hormones Department, National Research Centre, Dokki, Giza, Egypt
  • Chemistry Department, Faculty of Science, Helwan University, Cairo, Egypt
  • Chemistry Department, Faculty of Science, Helwan University, Cairo, Egypt
  • Chemistry of Tannins Department, National Research Centre, Dokki, Giza, Egypt
  • Hormones Department, National Research Centre, Dokki, Giza, Egypt
  • Applied Medical Sciences Department, Community College in Al-Qurayyat, Al-Jouf University, KSA
  • Chemistry Department, Faculty of Science, Helwan University, Cairo, Egypt
  • Ahmed HH, Shousha WGh, El-Mezayen HA, El-Toumy SA, Ramadan AR (2015) Updates on the biochemical and molecular mechanisms of N-nitrosodiethylamine induced hepatocellular carcinoma: Promising therapeutic role of Punica granatum peel Extract. Int J Pharm Sci Rev Res 32: 121-144.
  • Aleem E, Elshayeb A, Elhabachi N, Mansour AR, Gowily A, Hela A (2012) Serum IGFBP-3 is a more effective predictor than IGF-1 and IGF-2 for the development of hepatocellular carcinoma in patients with chronic HCV infection. Oncol Lett 3: 704-712. doi: 10.3892/ol.2011.546.
  • Armitage P, Berry G (1987) Comparison of several groups. In Statistical method in medical research, 2nd edn. pp 86-213. Oxford, Block well significant publication. MEDLINE
  • Barnes JM, Paget GE (1965) Mechanisms of toxic action. Prog Med Chem 4: 18-38.
  • Bio-Rad Laboratories Inc (2006) Real-time PCR applications guide. Bulletin 5279: 101.
  • Burr AW, Hillan KJ, Mclaughlin KE, Ferrier R, Chapman C, Mathew J, Burt AD (1996) Hepatocyte growth factor levels in liver and serum increase during chemical hepatocarcinogenesis. Hepatology 24: 1282-1287. doi: 10.1002/hep.510240549.
  • Capurro M, Wanless IR, Sherman M, Deboer G, Shi W, Miyoshi E, Filmus J (2003) Glypican-3: a novel serum and histochemical marker for hepatocellular carcinoma. Gastroenterology 125: 89-97. doi: 10.1016/S0016-5085(03)00689-9.
  • Capurro MI, Xiang YY, Lobe C, Filmus J (2005) Glypican-3 promotes the growth of hepatocellular carcinoma by stimulating canonical Wnt signaling. Cancer Res 65: 6245-6254. doi: 10.1158/0008-5472.CAN-04-4244.
  • Caruso ML, Valentini AM (1999) Overexpression of p53 in a large series of patients with hepatocellular carcinoma: a clinicopathological correlation. Anticancer Res 19: 3853-3856. doi: 10.1002/hep.1840160511.
  • Chan KL, Guan XY, Ng IO (2004) High-throughput tissue microarray analysis of c-myc activation inchronic liver diseases and hepatocellular carcinoma. Hum Pathol 35: 1324-1331. doi: 10.1016/j.humpath.2004.06.012.
  • Chang YF, Chi CW, Wang JJ (2006) Reactive oxygen species production is involved in quercetin-induced apoptosis in human hepatoma cells. Nutrition and Cancer 55: 201-209. doi: 10.1207/s15327914nc5502_12.
  • Chao JC, Chu CC (2004) Effects of Ginkgo biloba extract on cell proliferation and cytotoxicity in human hepatocellular carcinoma cells. World J Gastroenterol 10: 37-41. doi: 10.3748/wjg.v10.i1.37.
  • Chen G, Wang Y, Garate M, Zhou J, Li G (2010) The tumor suppressor ING3 is degraded by SCF (Skp2)-mediated ubiquitin-proteasome system. Oncogene 29: 1498-1508. doi: 10.1038/onc.2009.424.
  • Cohen GM (1997) Caspases: the executioners of apoptosis. Biochem J 326: 1-16.
  • Datta J, Kutay H, Nasser MW, Nuovo GJ, Wang B, Majumder S, Liu CG, Volinia S, Croce CM, Schmittgen TD, Ghoshal K, Jacob ST (2008) Methylation mediated silencing of MicroRNA-1 gene and its role in hepatocellular carcinogenesis. Cancer Res 68: 5049-5058. doi: 10.1158/0008-5472.CAN-07-6655.
  • Drury G, Wallington A (1980) Carleton's histological techniques. Oxford University Press: 403-406. MEDLINE
  • El-Sayyad HI, Ismail MF, Shalaby FM, Abou-El-Magd RF, Gaur RL, Fernando A, Raj MH, Ouhtit A (2009) Histopathological effects of cisplatin, doxorubicin and 5- flurouracil (5-Fu) on the liver of male albino rats. Int J Biol Sci 5: 466-473. doi: 10.7150/ijbs.5.466.
  • El-Zayadi AR, Badran HM, Barakat EM, Attia MD, Shawky S, Mohamed MK, Selim O, Saeid A (2005) Hepatocellular carcinoma in Egypt: a single center study over a decade. World J Gastroenterol 11: 5193-5198.
  • Eshak MG, Ghaly IS, Khalil WKB, Farag IM, Ghanem KZ (2010) Genetic alterations induced by toxic effect of thermally oxidized oil and protective role of tomatoes and carrots in mice. J Am Sci 6: 175-188. doi: 10.7537/marsjas060410.25
  • Farber E (1994) Programmed cell death: necrosis versus apoptosis. Mod Pathol 7: 605-609.
  • Farazi PA, DePinho RA (2006) Hepatocellular carcinoma pathogenesis: from genes to environment. Nat Rev Cancer 6: 674-687. doi: 10.1038/nrc1934.
  • Filmus J, Capurro M (2008) The role of glypican-3 in the regulation of body size and cancer. Cell Cycle 7: 2787-2790. doi: 10.4161/cc.7.18.6672.
  • Friesen C, Fulda S, Debatin KM (1999) Cytotoxic drugs and the CD95 pathway. Leukemia 13: 1854-1858. doi: 10.1038/sj.leu.2401333.
  • Fuke H, Shiraki K, Sugimoto K, Tanaka J, Beppu T, Yoneda K, Yamamoto N, Ito K, Masuya M, Takei Y (2007) Jak inhibitor induces S phase cell-cycle arrest and augments TRAIL-induced apoptosis in human hepatocellular carcinoma cells. Biochem Biophys Res Commun 363: 738-744.
  • Genoux E, Nicolle E, Boumendjel A (2011) Flavonoids as anticancer agents: recent progress and state of the art? Current Organic Chemistry 15: 2608-2615. doi: 10.2174/138527211796367363
  • Gupta C, Vikram A, Tripathi DN, Ramarao P, Jena GB (2010) Antioxidant and antimutagenic effect of quercetin against DEN induced hepatotoxicity in rat. Phytother Res 24: 119-128. doi: 10.1002/ptr.2883.
  • Harring TR, Kuten DA, Nguyen NT, Goss JA, O'Mahony CA (2011) Orthotopic liver transplantation in patients with mixed hepatocellular carcinoma - cholangiocarcinoma. J Transplant Technol Res 1: 104. doi: 10.4172/2161-0991.1000104
  • He TC, Sparks AB, Rago C, Hermeking H, Zawel L, da Costa LT, Morin PJ, Vogelstein B, Kinzler KW (1998) Identification of c-MYC as a target of the APC pathway. Science 281: 1509-1512. doi: 10.1126/science.281.5382.1509.
  • Herrera B, Fernandez M, Alvarez A, Roncero C, Benito M, Gil J, Fabregat I (2001) Activation of caspases occurs downstream from radical oxygen species production, Bcl-xL downregulation, and early cytochrome C release in apoptosis induced by transforming growth factor beta in rat fetal hepatocytes. Hepatology 34: 548-556. doi: 10.1053/jhep.2001.27447.
  • Jose K, Kuttan R, Bhattacharya K (1998) Effect of Emblica officinalis extract on hepatocarcinogenesis and carcinogen metabolism. J Clin Biochem Nutr 25: 31-39. doi: 10.3164/jcbn.25.31
  • Karimov KH, Inoyatova FK, Mukhamedova MT (2003) Changes in some indices of the synthesis of nitric oxide during the early stages of hepatocarcinogenesis. Exp Toxicol Pathol 55: 17-19. doi: 10.1078/0940-2993-00267.
  • Kawakami K, Enokida H, Chiyomaru T, Tatarano S, Yoshino H, Kagara I, Gotanda T, Tachiwada T, Nishiyama K, Nohata N, Seki N, Nakagawa M (2012) The functional significance of miR-1 and miR-133a in renal cell carcinoma. Eur J Cancer 48: 827-836. doi: 10.1016/j.ejca.2011.06.030.
  • Kim TH, Zhao Y, Barber MJ, Kuharsky DK, Yin XM (2000) Bid-induced cytochrome c release is mediated by a pathway independent of mitochondrial permeability transition pore and Bax. J Biol Chem 275: 39474-39481. doi: 10.1074/jbc.M003370200.
  • Koon HB, Ippolito GC, Banham AH, Tucker PW (2007) Foxp1: a potential therapeutic target in cancer. Expert Opin Ther Targets 11: 955-965. doi: 10.1517/14728222.11.7.955.
  • Lazarevich NL (2000) Molecular mechanisms of alpha-fetoprotein gene expression. Biochemistry (Mosc.) 65: 117-133.
  • Lee WJ, Shim JY, Zhu BT (2005) Mechanisms for the inhibition of DNA methyltransferases by tea catechins and bioflavonoids. Mol Pharmacol 68: 1018-1030. doi: 10.1124/mol.104.008367.
  • Lehman EM, Soliman AS, Ismail K, Hablas A, Seifeldin IA, Ramadan M, El-Hamzawy H, Shoushtari CS, Wilson ML (2008) Patterns of hepatocellular carcinoma incidence in Egypt from a population-based Cancer registry. Hepatol Res 38: 465-473. doi: 10.1111/j.1872-034X.2007.00299.x.
  • Li L, Jin R, Zhang X, Lv F, Liu L, Liu D, Liu K, Li N, Chen D (2012a) Oncogenic activation of glypican-3 by c-Myc in human hepatocellular carcinoma. Hepatology 56: 1380-1390. doi: 10.1002/hep.25891.
  • Li D, Yang P, Li H, Cheng P, Zhang L, Wei D, Su X, Peng J, Gao H, Tan Y, Zhao Z, Li Y, Qi Z, Rui Y, Zhang T (2012b) MicroRNA-1 inhibits proliferation of hepatocarcinoma cells by targeting endothelin-1. Life Sci 91: 440-447. doi: 10.1016/j.lfs.2012.08.015.
  • Liao DJ, Blanck A, Eneroth P, Gustafsson JA, Hällström IP (2001) Diethylnitrosamine causes pituitary damage, disturbs hormone levels, and reduces sexual dimorphism of certain liver functions in the rat. Environ Health Perspect 109: 943-947.
  • Lu M, Chen F, Wang Q, Wang K, Pan Q, Zhang X (2012) Downregulation of inhibitor of growth 3 is correlated with tumorigenesis and progression of hepatocellular carcinoma. Oncology Letters 4: 47-52. doi: 10.3892/ol.2012.685.
  • Łubgan D, Marczak A, Walczak M, Distel L, Jóźwiak Z (2006) Pharmacological mechanisms of doxorubicin activity (DOX) - current state of knowledge. Przegl Lek 63: 782–788.
  • Luo J, Shah S, Riabowol K, Mains PE (2009) The Caenorhabditis elegans ING-3 gene regulates ionizing radiation-induced germ-cell apoptosis in a p53-associated pathway. Genetics 181: 473-482. doi: 10.1534/genetics.107.080515.
  • Macnab GM, Urbanowicz JM, Kew MC (1978) Carcinoembryonic antigen in hepatocellular cancer. Br J Cancer 38: 51-54. doi: 10.1038/bjc.1978.162.
  • Marcocci L, Packer L, Droy-Lefaix MT, Sekaki A, Gardès-Albert M (1994) Antioxidant action of Ginkgo biloba extract EGb 761. Methods Enzymol 234: 462-475.
  • Middleton E Jr, Kandaswami C, Theoharides TC (2000) The effects of plant flavonoids on mammalian biology: implications for immunity, inflammation and cancer. Pharmacol Rev 52: 673-751. doi: 10.1007/978-1-4899-2911-2_15.
  • Motalleb G, Hanachi P, Fauziah O, Asmah R (2008) Effect of Berberis vulgaris fruit extract on alpha-fetoprotein gene expression and chemical carcinogen metabolizing enzymes activities in hepatocarcinogenesis rats. Iranian J Cancer Prevention 1: 33-44.
  • Nagashima M, Shiseki M, Pedeux RM, Okamura S, Kitahama-Shiseki M, Miura K, Yokota J, Harris CC (2003) A novel PHD-finger motif protein, p47ING3, modulates p53-mediated transcription, cell cycle control and apoptosis. Oncogene 22: 343-350. doi: 10.1038/sj.onc.1206115.
  • Nakamura T, Ueda Y, Juan Y, Katsuda S, Takahashi H, Koh E (2000) Fas-mediated apoptosis in adriamycin induced cardiomyopathy in rats: in vivo study. Circulation 102: 572-578. doi: 10.1161/01.CIR.102.5.572.
  • Ogden SK, Lee KC, Wernke-Dollries K, Stratton SA, Aronow B, Barton MC (2001) p53 targets chromatin structure alteration to repress alpha-fetoprotein gene expression. J Biol Chem 276: 42057-42062. doi: 10.1074/jbc.C100381200.
  • Oka H, Saito A, Ito K, Kumada T, Satomura S, Kasugai H, Osaki Y, Seki T, Kudo M, Tanaka M (2001) Multicenter prospective analysis of newly diagnosed hepatocellular carcinoma with respect to the percentage of Lens culinaris agglutinin-reactive alpha-fetoprotein. J Gastroenterol Hepatol 16: 1378-1383. doi: 10.1046/j.1440-1746.2001.02643.x.
  • Pan G, Ni J, Wei YF, Yu G, Gentz R, Dixit VM (1997) An antagonist decoy receptor and a death domain-containing receptor for TRAIL. Science 277: 815. doi: 10.1126/science.277.5327.815.
  • Park CH, Chang JY, Hahm ER, Park S, Kim HK, Yang CH (2005) Quercetin, a potent inhibitor against b-catenin/Tcf signaling in SW480 colon cancer cells. Biochem Biophys Res Commun 328: 227-234. doi: 10.1016/j.bbrc.2004.12.151.
  • Pratt WB, Ruddon RW, Ensminger WD, Maybaum J (1994) The anticancer drugs. Oxford University Press. New York. MEDLINE
  • Premalatha B, Sachdanandam P (1999) Effects of semecarpus anacardium nut milk extract on rat serum alpha-fetoprotein level in aflatoxin B1-mediated hepatocellular carcinoma. Fitoterapia 70: 279-283. doi: 10.1016/S0367-326X(99)00043-X
  • Puttalingamma V (2015) Ginkgo biloba living fossil, wonderful medicinal plant - a review. Int J Adv Res 3: 506–511.
  • Ramakrishnan G, Jagan S, Kamaraj S, Anandakumar P, Devaki T (2009) Silymarin attenuated mast cell recruitment thereby decreased the expressions of matrix metalloproteinases-2 and 9 in rat liver carcinogenesis. Invest New Drugs 27: 233-240. doi: 10.1007/s10637-008-9163-y.
  • Reid JF, Sokolova V, Zoni E, Lampis A, Pizzamiglio S, Bertan C, Zanutto S, Perrone F, Camerini T, Gallino G, Verderio P, Leo E, Pilotti S, Gariboldi M, Pierotti MA (2012) miRNA profiling in colorectal cancer highlights miR-1 involvement in MET-dependent proliferation. Mol Cancer Res 10: 504-515. doi: 10.1158/1541-7786.MCR-11-0342.
  • Russo M, Spagnuolo C, Tedesco I, Bilotto S, Russo GL (2012) The flavonoid quercetin in disease prevention and therapy: facts and fancies. Biochemical Pharmacology 83: 6-15. doi: 10.1016/j.bcp.2011.08.010.
  • Saad SY, Najja TA, Al-Rikabi AC (2001) The preventive role of deferoxamine against acute doxorubicin-induced cardiac, renal and hepatic toxicity in rats. Pharmacol Res 43: 211-218. doi: 10.1006/phrs.2000.0769.
  • Sadiknah NAH, EL-Maraghy SA, Ismail MF (2008) Diethylnitrosamine induced hepatocarcino-genesis in rats: possible chemoprevention by blueberries. African J Bioch Res 2: 081-087.
  • Sagar SM, Yance D, Wong RK (2006) Natural health products that inhibit angiogenesis: a potential source for investigational new agents to treat cancer-Part 1. Curr Oncol 13: 14-26.
  • Sapisochin G, Charco R (2011) Salvage liver transplantation for HCC: an old story without consensus? J Transplant Technol Res 1: 06. doi: 10.4172/2161-0991.1000106e
  • Scherer E, Emmelot P (1975) Kinetics of induction and growth of precancerous liver cell foci, and liver tumour formation by diethylnitrosamine in the rat. Eur J Cancer 11: 689-696. doi: 10.1016/0014-2964(75)90042-0.
  • Shu W, Lu MM, Zhang Y, Tucker PW, Zhou D, Morrisey EE (2007) Foxp2 and Foxp1 cooperatively regulate lung and esophagus development. Development 134: 1991-2000. doi: 10.1242/dev.02846.
  • Sivaramakrishnan V, Shilpa PN, Praveen Kumar VR, Niranjali Devaraj S (2008) Attenuation of N-nitrosodiethylamine-induced hepatocellular carcinoma by a novel flavonol-Morin. Chem Biol Interact 171: 79-88. doi: 10.1016/j.cbi.2007.09.003.
  • Suzuki MI, Sugimoto K, Tanaka J, Tameda M, Inagaki Y, Kusagawa S, Nojiri K, Beppu T, Yoneda K, Yamamoto N, Ito M, Yoneda M, Uchida K, Takase K, Shiraki K (2010) Up-regulation of glypican-3 in human hepatocellular carcinoma. Anticancer Res 30: 5055-5061.
  • Tanigawa S, Fujii M, Hou DX (2008) Stabilization of p53 is involved in quercetin-induced cell cycle arrest and apoptosis in HepG2 cells. Biosci Biotechnol Biochem 72: 797-804. doi: 10.1271/bbb.70680.
  • Thomas P, Hems DA (1975) The hepatic clearance of circulating native and asialo carcinoembryonic antigen by the rat. Biochem Biophys Res Comm 67: 1205-1209.
  • Van Beek TA (2002) Chemical analysis of Ginkgo biloba leaves and extracts. J Chromatogr A 967: 21-55. doi: 10.1016/j.chroma.2009.01.013.
  • Verna L, Whysner J, Williams GM (1996) N-Nitrosodiethylamine mechanistic data and risk assessment: bioactivation, DNA-adduct formation, mutagenicity and tumor initiation. Pharmacol Ther 71: 57-81. doi: 10.1016/0163-7258(96)00062-9.
  • Wang Y, Li G (2006) ING3 promotes UV-induced apoptosis via Fas/caspase-8 pathway in melanoma cells. J Biol Chem 281: 11887-11893. doi: 10.1074/jbc.M511309200.
  • Wang Y, Shen Y (2013) Unresectable hepatocellular carcinoma treated with transarterial chemoembolization: clinical data from a single teaching hospital. Int J Clin Exp Med 6: 367-371.
  • Yang CS, Fang M, Lambert JD, Yan P, Huang TH (2008) Reversal of hypermethylation and reactivation of genes by dietary polyphenolic compounds. Nutr Rev 66: S18-S20. doi: 10.1111/j.1753-4887.2008.00059.x.
  • Yang HY, Liu HL, Tian LT, Song RP, Song X, Yin DL, Liang YJ, Qu LD, Jiang HC, Liu JR, Liu LX (2012) Expression and prognostic value of ING3 in human primary hepatocellular carcinoma. Experimental Biology and Medicine 237: 352-361. doi: 10.1258/ebm.2011.011346.
  • Yokochi T, Robertson KD (2004) Doxorubicin inhibits DNMT1, resulting in conditional apoptosis. Mol Pharmacol 66: 1415-1420. doi: 10.1124/mol.104.002634.
  • Zhang XL, Yu H, Xiong YY, Ma ST, Zhao L, She SF (2013) Resveratrol down-regulates myosin light chain kinase, induces apoptosis and inhibits diethylnitrosamine-induced liver tumorigenesis in rats. Int J Mol Sci 14: 1940-1951. doi: 10.3390/ijms14011940.
  • Zhang Y, Zhang S, Wang X, Liu J, Yang L, He S, Chen L, Huang J (2012) Prognostic significance of Foxp1 as an oncogene in hepatocellular carcinoma. J Clin Pathol 65: 528-533. doi: 10.1136/jclinpath-2011-200547.
  • Zhou L, Liu J, Luo F (2006) Serum tumor markers for detection of hepatocellular carcinoma. World J Gastroenterol 12: 1175-1181. doi: 10.3748/wjg.v12.i8.1175.
  • Zimmer R, Thomas P (2001) Mutations in the carcinoembryonic antigen gene in colorectal cancer patients: implications on liver metastasis. Cancer Res 61: 2822-2826.
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