Effect of geraniol and vitamin C on diethylnitrosamine induced experimental hepatocellular carcinogenesis
Year 2022,
Volume: 47 Issue: 2, 738 - 746, 30.06.2022
Gamze Taşkın Şenol
,
Halil Mahir Kaplan
,
Neslihan Boyan
,
Özkan Oğuz
,
Ergin Şingirik
Abstract
Purpose: This study aimed to investigate the protective effect of geraniol and vitamin C on the experimental hepatocellular carcinogenesis (HCC) model by inducing FL83B hepatocyte cell lines with diethylnitrosamine (DENA).
Materials and Methods: The cells prepared in the medium were incubated with DENA (5 μM), geraniol (5 μM), and vitamin C (50 μM) for 48 hours in an incubator at 37 °C and 5% CO2. Groups were designed as follows: Group 1 (Control), group 2 (DENA Control), group 3 (DENA+Geraniol), group 4 (DENA+Vitamin C), and group 5 (DENA+Geraniol+Vitamin C) on standard cell culture plates. Six plates from each experimental group were studied. After the homogenization was centrifuged, analyses of pathway mediators NF-ĸB, AIF, caspase-3, BCL-2, bax, gadd153, GRP78, and COX were performed by the Elisa method.
Results: The expression of Bax, caspase-3, COX-2, NFkB, GADD153, AIF, and GRP78 increased in cancer cells when compared to group 1 and decreased in other groups where antiproliferative agents were applied. Bcl-2 expression is decreased when compared to group 1, and expression is increased in other groups where antiproliferative agents are applied.
Conclusion: There was a significant hepatoprotective effect in the groups administered geraniol+vitamin C on pathway mediators in a DENA-induced HCC model.
Supporting Institution
Çukurova Üniversitesi Bilimsel Araştırmalar Projeleri Bİrimi
Project Number
TDK-2017-9717
Thanks
The authors thanks to Ahmet Hilmi Yücel, Prof. Dr., who suddenly passed away and had a huge contribution to the study that is in the working phase and thanks for the using laboratory facilities to the Cukurova University Department of Medical Pharmacology.
References
- Kurma K, Manches O, Chuffart F, Sturm N, Gharzeddine K, Zhang J, et al. DEN-induced rat model reproduces key features of human hepatocellular carcinoma. Cancers. 2021;13(19):1–15. DOI: 10.3390/cancers13194981.
- Song J, Ding W, Liu B, Liu D, Xia Z, Zhang L, et al. Anticancer effect of caudatin in diethylnitrosamine-induced hepatocarcinogenesis in rats. Molecular Medicine Reports. 2020;22(2):697–706. DOI: 10.3892/mmr.2020.11135.
- Karakurt E, Dağ S. Hepatoprotective effect of silymarin against diethylnitrosamine induced oxidative-nitrosative stress in rats. Van Veterinary Journal. 2020;31(2):105–10. DOI: 10.36483/vanvetj.680365.
- You Y, Zhu F, Li Z, Zhang LF, Xie Y, Chinnathambi A, et al. Phyllanthin prevents diethylnitrosamine (DEN) induced liver carcinogenesis in rats and induces apoptotic cell death in HepG2 cells. Biomedicine and Pharmacotherapy. 2021;137(January):111335. DOI: 10.1016/j.biopha.2021.111335.
- Liu K, Chen J, McCaughan GW. Animal models for hepatocellular carcinoma arising from alcoholic and metabolic liver diseases. Hepatoma Research. 2020;2020. DOI: 10.20517/2394-5079.2019.39.
- Zhi H, Cui J, Yang H, Zhang Y, Zhu M. Research progress of geraniol in tumor therapy. Proceedings of Anticancer Research. 2021;5(1):2–6. DOI: 10.26689/par.v5i1.1882.
- el Azab EF, Elguindy NM, Yacout GA, Elgamal DA. Hepatoprotective impact of geraniol against ccl4-induced liver fibrosis in rats. Pakistan Journal of Biological Sciences. 2020;23(12):1650–8. DOI. 10.3923/pjbs.2020.1650.1658.
- Mohammed MJ, Tadros MG, Michel HE. Geraniol protects against cyclophosphamide-induced hepatotoxicity in rats: Possible role of MAPK and PPAR-γ signaling pathways. Food and Chemical Toxicology. 2020;139. DOI: 10.1016/j.fct.2020.111251.
- Blowman K, Magalhães M, Lemos MFL, Cabral C, Pires IM. Anticancer properties of essential oils and other natural products. Evidence-based Complementary and Alternative Medicine. 2018;(3149362):1–12. DOI: 10.1155/2018/3149362.
- Queiroz TB, Santos GF, Ventura SC, Hiruma-Lima CA, Gaivão IOM, Maistro EL. Cytotoxic and genotoxic potential of geraniol in peripheral blood mononuclear cells and human hepatoma cell line (HepG2). Genetics and Molecular Research. 2017;16(3). DOI: 10.4238/gmr16039777.
- Maczka W, Winska K, Grabarczyk M. One hundred faces of geraniol. Molecules. 2020;25(14):1–16. DOI: 10.3390/molecules25143303.
- Satheesh NJ, Samuel SM, Büsselberg D. Combination therapy with vitamin C could eradicate cancer stem cells. Biomolecules. 2020;10(1). DOI: 10.3390/biom10010079.
- Alexandrova R, Gavrilov I. Cell cultures as model systems in breast cancer research. Article in Merit Research Journal of Medicine and Medical Sciences. 2019;(February). Available from: https://www.researchgate.net/publication/333185614.
- Ortiz N, Jiménez MF, Chaverri C, Cicció JF, Díaz C. Effect on cell growth, viability and migration of geraniol and geraniol-containing essential oil from Lippia alba (Verbenaceae) on gastric carcinoma cells. Journal of Essential Oil Research. 2021;00(00):1–12. DOI: 10.1080/10412905.2021.1975576.
- Crespo R, Rodenak-Kladniew BE, Castro MA, Soberón M v., Lavarías SML. Induction of oxidative stress as a possible mechanism by which geraniol affects the proliferation of human A549 and HepG2 tumor cells. Chemico-Biological Interactions. 2020;320(February):109029. DOI: 10.1016/j.cbi.2020.109029.
- Rahmouni F, Badraoui R, Amri N, Elleuch A, El-Feki A, Rebai T, et al. Hepatotoxicity and nephrotoxicity in rats induced by carbon tetrachloride and the protective effects of Teucrium polium and vitamin C. Toxicology Mechanisms and Methods. 2019;29(5):313–21. DOI: 10.1080/15376516.2018.1519864.
- Grosso G, Bei R, Mistretta A, Marventano S, Calabrese G, Masuelli L, et al. Effects of vitamin C on health: A review of evidence. Frontiers in Bioscience. 2013;18(3):1017–29. DOI: 10.2741/4160.
- Al-Hrout A, Chaiboonchoe A, Khraiwesh B, Murali C, Baig B, El-Awady R, et al. Safranal induces DNA double-strand breakage and ER-stress-mediated cell death in hepatocellular carcinoma cells. Scientific Reports. 2018;8(1):1–15. DOI: 10.1038/s41598-018-34855-0.
- Lin X-H, Qiu B-Q, Ma M, Zhang R, Hsu S-J, Liu H-H, et al. Suppressing DRP1-mediated mitochondrialfissionand mitophagy increases mitochondrial apoptosisof hepatocellular carcinoma cells in the setting ofhypoxia. Oncogenesis. 2020;9(67):1–13. DOI: 10.1038/s41389-020-00251-5.
- Wang C, Gao D, Guo K, Kang X, Jiang K, Sun C, et al. Novel synergistic antitumor effects of rapamycin with bortezomib on hepatocellular carcinoma cells and orthotopic tumor model. BMC Cancer. 2012;12(1):166. DOI: 10.1186/1471-2407-12-166.
- Dasmahapatra G, Yerram N, Dai Y, Dent P, Grant S. Synergistic interactions between vorinostat and sorafenib in chronic myelogenous leukemia cells involve Mcl-1 and p21CIP1 down-regulation. Clinical Cancer Research. 2007;13(14):4280–90. DOI: 10.1158/1078-0432.CCR-07-0835.
- Shen CH, Tung SY, Huang WS, Lee KF, Hsieh YY, Hsieh MC, et al. Comparative proteomic identification of protein disulphide isomerase A6 associated with tert-butylhydroperoxide-induced liver injury in rat hepatocytes. Cellular Physiology and Biochemistry. 2018;45(5):1915–26. DOI: 10.1159/000487968.
- Prasanna PL, Renu K, Valsala Gopalakrishnan A. New molecular and biochemical insights of doxorubicin-induced hepatotoxicity. Life Sciences. 2020;250(February):117599. DOI: 10.1016/j.lfs.2020.117599.
- Abdulrazzaq AM, Badr M, Gammoh O, Abu Khalil AA, Ghanim BY, Alhussainy TM, et al. Hepatoprotective actions of ascorbic acid, alpha lipoic acid and silymarin or their combination against acetaminophen-induced hepatotoxicity in rats. Medicina (Lithuania). 2019;55(5). DOI: 10.3390/medicina55050181.
- Abdulkhaleq FM, Alhussainy TM, Badr MM, Abu Khalil AA, Gammoh O, Ghanim BY, et al. Antioxidative stress effects of vitamins C, E, and B 12 , and their combination can protect the liver against acetaminophen-induced hepatotoxicity in rats. Drug Design, Development and Therapy. 2018;12:3525–33. DOI: 10.2147/DDDT.S172487.
- Komeil IA, El-Refaie WM, Gowayed MA, El-Ganainy SO, el Achy SN, Huttunen KM, et al. Oral genistein-loaded phytosomes with enhanced hepatic uptake, residence and improved therapeutic efficacy against hepatocellular carcinoma. International Journal of Pharmaceutics. 2021;601(November 2020):120564. DOI: 10.1016/j.ijpharm.2021.120564.
- Elkeiy MM, Khamis AA, El-Gamal MM, Abo Gazia MM, Zalat ZA, El-Magd MA. Chitosan nanoparticles from Artemia salina inhibit progression of hepatocellular carcinoma in vitro and in vivo. Environmental Science and Pollution Research. 2020;27(16):19016–28. DOI: 10.1007/s11356-018-3339-6.
Geraniol ve vitamin C’nin dietilnitrozamin kaynaklı deneysel hepatoselüler karsinogenez üzerindeki etkisi
Year 2022,
Volume: 47 Issue: 2, 738 - 746, 30.06.2022
Gamze Taşkın Şenol
,
Halil Mahir Kaplan
,
Neslihan Boyan
,
Özkan Oğuz
,
Ergin Şingirik
Abstract
Amaç: Bu çalışmanın amacı dietilnitrozamin (DENA) ile FL83B hepatosit hücre hattını indükleyerek oluşturulan deneysel hepatosellüler karsinogenezis (HSK) modelinde geraniol ve vitamin C'nin koruyucu etkisini araştırmaktır.
Gereç ve Yöntem: Bu amaçla çalışmamızda besiyerinde hazırlanan hücreler DENA (5 μM), geraniol (5 μM) ve vitamin C (50 μM) ile 37 °C sıcaklıkta ve %5’lik CO2‘li etüvde 48 saat inkübe edildi. Hücreler inkübe edildikten sonra gruplar standart hücre kültürü plakalarında Grup 1 (Kontrol), grup 2 (DENA Kontrol), grup 3 (DENA+Geraniol), grup 4 (DENA+Vitamin C) ve grup 5 (DENA+Geraniol+Vitamin C) olmak üzere beş gruba ayrıldı. Her deney grubundan altı plaka çalışıldı. Homojenatlar santrifüj edildikten sonra yolak mediyatörlerinden NF-ĸB, AIF, caspase-3, BCL-2, bax, gadd153, GRP78, and COX-2 analizleri Elisa metodu ile yapıldı.
Bulgular: Kanser hücrelerinde Bax, caspase-3, COX-2, NFkB, GADD153, AIF ve GRP78 ekspresyonu grup 1 ile kıyaslandığında artmakta, geraniol ve vitamin C’nin uygulandığı diğer gruplarda ekspresyonun azalmakta olduğu belirlenmiştir. Bcl-2 ekspresyonu grup 1 ile kıyaslandığında azalmakta, geraniol ve vitamin C’nin uygulandığı diğer gruplarda ekspresyon artmaktadır.
Sonuç: Bu çalışmada, DENA ile indüklenen HCC modelinde geraniol ve vitamin C uygulanan gruplarda yolak mediyatörleri aracılığıyla önemli bir hepatoprotektif etki olduğu gözlendi.
Project Number
TDK-2017-9717
References
- Kurma K, Manches O, Chuffart F, Sturm N, Gharzeddine K, Zhang J, et al. DEN-induced rat model reproduces key features of human hepatocellular carcinoma. Cancers. 2021;13(19):1–15. DOI: 10.3390/cancers13194981.
- Song J, Ding W, Liu B, Liu D, Xia Z, Zhang L, et al. Anticancer effect of caudatin in diethylnitrosamine-induced hepatocarcinogenesis in rats. Molecular Medicine Reports. 2020;22(2):697–706. DOI: 10.3892/mmr.2020.11135.
- Karakurt E, Dağ S. Hepatoprotective effect of silymarin against diethylnitrosamine induced oxidative-nitrosative stress in rats. Van Veterinary Journal. 2020;31(2):105–10. DOI: 10.36483/vanvetj.680365.
- You Y, Zhu F, Li Z, Zhang LF, Xie Y, Chinnathambi A, et al. Phyllanthin prevents diethylnitrosamine (DEN) induced liver carcinogenesis in rats and induces apoptotic cell death in HepG2 cells. Biomedicine and Pharmacotherapy. 2021;137(January):111335. DOI: 10.1016/j.biopha.2021.111335.
- Liu K, Chen J, McCaughan GW. Animal models for hepatocellular carcinoma arising from alcoholic and metabolic liver diseases. Hepatoma Research. 2020;2020. DOI: 10.20517/2394-5079.2019.39.
- Zhi H, Cui J, Yang H, Zhang Y, Zhu M. Research progress of geraniol in tumor therapy. Proceedings of Anticancer Research. 2021;5(1):2–6. DOI: 10.26689/par.v5i1.1882.
- el Azab EF, Elguindy NM, Yacout GA, Elgamal DA. Hepatoprotective impact of geraniol against ccl4-induced liver fibrosis in rats. Pakistan Journal of Biological Sciences. 2020;23(12):1650–8. DOI. 10.3923/pjbs.2020.1650.1658.
- Mohammed MJ, Tadros MG, Michel HE. Geraniol protects against cyclophosphamide-induced hepatotoxicity in rats: Possible role of MAPK and PPAR-γ signaling pathways. Food and Chemical Toxicology. 2020;139. DOI: 10.1016/j.fct.2020.111251.
- Blowman K, Magalhães M, Lemos MFL, Cabral C, Pires IM. Anticancer properties of essential oils and other natural products. Evidence-based Complementary and Alternative Medicine. 2018;(3149362):1–12. DOI: 10.1155/2018/3149362.
- Queiroz TB, Santos GF, Ventura SC, Hiruma-Lima CA, Gaivão IOM, Maistro EL. Cytotoxic and genotoxic potential of geraniol in peripheral blood mononuclear cells and human hepatoma cell line (HepG2). Genetics and Molecular Research. 2017;16(3). DOI: 10.4238/gmr16039777.
- Maczka W, Winska K, Grabarczyk M. One hundred faces of geraniol. Molecules. 2020;25(14):1–16. DOI: 10.3390/molecules25143303.
- Satheesh NJ, Samuel SM, Büsselberg D. Combination therapy with vitamin C could eradicate cancer stem cells. Biomolecules. 2020;10(1). DOI: 10.3390/biom10010079.
- Alexandrova R, Gavrilov I. Cell cultures as model systems in breast cancer research. Article in Merit Research Journal of Medicine and Medical Sciences. 2019;(February). Available from: https://www.researchgate.net/publication/333185614.
- Ortiz N, Jiménez MF, Chaverri C, Cicció JF, Díaz C. Effect on cell growth, viability and migration of geraniol and geraniol-containing essential oil from Lippia alba (Verbenaceae) on gastric carcinoma cells. Journal of Essential Oil Research. 2021;00(00):1–12. DOI: 10.1080/10412905.2021.1975576.
- Crespo R, Rodenak-Kladniew BE, Castro MA, Soberón M v., Lavarías SML. Induction of oxidative stress as a possible mechanism by which geraniol affects the proliferation of human A549 and HepG2 tumor cells. Chemico-Biological Interactions. 2020;320(February):109029. DOI: 10.1016/j.cbi.2020.109029.
- Rahmouni F, Badraoui R, Amri N, Elleuch A, El-Feki A, Rebai T, et al. Hepatotoxicity and nephrotoxicity in rats induced by carbon tetrachloride and the protective effects of Teucrium polium and vitamin C. Toxicology Mechanisms and Methods. 2019;29(5):313–21. DOI: 10.1080/15376516.2018.1519864.
- Grosso G, Bei R, Mistretta A, Marventano S, Calabrese G, Masuelli L, et al. Effects of vitamin C on health: A review of evidence. Frontiers in Bioscience. 2013;18(3):1017–29. DOI: 10.2741/4160.
- Al-Hrout A, Chaiboonchoe A, Khraiwesh B, Murali C, Baig B, El-Awady R, et al. Safranal induces DNA double-strand breakage and ER-stress-mediated cell death in hepatocellular carcinoma cells. Scientific Reports. 2018;8(1):1–15. DOI: 10.1038/s41598-018-34855-0.
- Lin X-H, Qiu B-Q, Ma M, Zhang R, Hsu S-J, Liu H-H, et al. Suppressing DRP1-mediated mitochondrialfissionand mitophagy increases mitochondrial apoptosisof hepatocellular carcinoma cells in the setting ofhypoxia. Oncogenesis. 2020;9(67):1–13. DOI: 10.1038/s41389-020-00251-5.
- Wang C, Gao D, Guo K, Kang X, Jiang K, Sun C, et al. Novel synergistic antitumor effects of rapamycin with bortezomib on hepatocellular carcinoma cells and orthotopic tumor model. BMC Cancer. 2012;12(1):166. DOI: 10.1186/1471-2407-12-166.
- Dasmahapatra G, Yerram N, Dai Y, Dent P, Grant S. Synergistic interactions between vorinostat and sorafenib in chronic myelogenous leukemia cells involve Mcl-1 and p21CIP1 down-regulation. Clinical Cancer Research. 2007;13(14):4280–90. DOI: 10.1158/1078-0432.CCR-07-0835.
- Shen CH, Tung SY, Huang WS, Lee KF, Hsieh YY, Hsieh MC, et al. Comparative proteomic identification of protein disulphide isomerase A6 associated with tert-butylhydroperoxide-induced liver injury in rat hepatocytes. Cellular Physiology and Biochemistry. 2018;45(5):1915–26. DOI: 10.1159/000487968.
- Prasanna PL, Renu K, Valsala Gopalakrishnan A. New molecular and biochemical insights of doxorubicin-induced hepatotoxicity. Life Sciences. 2020;250(February):117599. DOI: 10.1016/j.lfs.2020.117599.
- Abdulrazzaq AM, Badr M, Gammoh O, Abu Khalil AA, Ghanim BY, Alhussainy TM, et al. Hepatoprotective actions of ascorbic acid, alpha lipoic acid and silymarin or their combination against acetaminophen-induced hepatotoxicity in rats. Medicina (Lithuania). 2019;55(5). DOI: 10.3390/medicina55050181.
- Abdulkhaleq FM, Alhussainy TM, Badr MM, Abu Khalil AA, Gammoh O, Ghanim BY, et al. Antioxidative stress effects of vitamins C, E, and B 12 , and their combination can protect the liver against acetaminophen-induced hepatotoxicity in rats. Drug Design, Development and Therapy. 2018;12:3525–33. DOI: 10.2147/DDDT.S172487.
- Komeil IA, El-Refaie WM, Gowayed MA, El-Ganainy SO, el Achy SN, Huttunen KM, et al. Oral genistein-loaded phytosomes with enhanced hepatic uptake, residence and improved therapeutic efficacy against hepatocellular carcinoma. International Journal of Pharmaceutics. 2021;601(November 2020):120564. DOI: 10.1016/j.ijpharm.2021.120564.
- Elkeiy MM, Khamis AA, El-Gamal MM, Abo Gazia MM, Zalat ZA, El-Magd MA. Chitosan nanoparticles from Artemia salina inhibit progression of hepatocellular carcinoma in vitro and in vivo. Environmental Science and Pollution Research. 2020;27(16):19016–28. DOI: 10.1007/s11356-018-3339-6.