Sains Malaysiana 48(2)(2019): 347–352

http://dx.doi.org/10.17576/jsm-2019-4802-11

The Effects of Oleuropein on Apoptotic Rate and Oxidative Stress Profiles during Tumour Promotion Stage in the Mouse Skin Carcinogenesis Model

(Kesan Oleuropein atas Kadar Apoptosis dan Tekanan Oksidatif Profil semasa Peringkat Galakan Tumor dalam Model Karsinogenesis Kulit Tikus)

SITI FATHIAH MASRE*, AZIM IZZUDDEEN, DAYANG NOOR SUZLIANA JOHN & ZARIYANTEY ABDUL HAMID

Biomedical Science Programme, Centre of Health and Applied Sciences, Faculty of Health Sciences, Universiti Kebangsaan Malaysia, Jalan Raja Muda Aziz, 50300 Kuala Lumpur, Federal Territory, Malaysia

Diserahkan: 19 Februari 2018/Diterima: 5 Jun 2018

ABSTRACT

Oleuropein is a phenolic compound that can be abundantly found in the olive plant and it possesses pharmacological properties including anticancer, antioxidant, and anti-inflammatory. This present study was designed to determine the effects of oleuropein on tumour promotion stage, particularly on the histopathological changes, apoptotic rates and oxidative stress profiles by using the mouse skin carcinogenesis model. Female ICR mice were randomly divided into 3 groups (n= 8 mice per group) as follows: Control induced with DMBA/TPA, negative control (acetone) and oleuropein-treated groups. For the treatment group, the mice were initiated with DMBA (200 nmol) followed by pre-treatment with oleuropein (10 mg/kg) and subsequent promotion with TPA (20 nmol). The treatments were topically applied on the shaved dorsal up to 10 weeks. Histopathology analysis showed that oleuropein-pretreated group appeared lack of thickness in epidermal hyperplasia, as compared to thick hyperplasia and epidermal disorganisation in the DMBA/TPA control group. Data also showed that oleuropein pre-treatment resulted in a significant increase of the apoptotic rates (p<0.05) as indicated by the activated caspase-3 labelling compared to DMBA/TPA group. Interestingly, the level of MDA is significantly reduced (p<0.05) in the oleuropein pre-treated group compared to DMBA/TPA group. Next, pre-treatment of oleuropein caused a significant decrease in the GSH levels (p<0.05) along with a significant increase in the SOD levels (p<0.05) compared to the DMBA/TPA group. Overall, this study indicates that oleuropein may act as a potential chemopreventive agent through its apoptotic and antioxidant defence activities on tumour promotion stage in skin carcinogenesis event.

Keywords: Carcinogenesis; chemoprevention; oleuropein; skin cancer; tumour promotion

ABSTRAK

Oleuropein adalah sebatian fenolik yang banyak ditemui dalam pokok zaitun dan ia mempunyai beberapa aktiviti farmakologi termasuk antikanser, antioksidan dan anti-inflamasi. Kajian ini dijalankan untuk menentukan kesan oleuropein pada peringkat galakan tumor terhadap perubahan histopatologi, kadar apoptosis dan profil tekanan oksidatif dalam model karsinogenesis kulit tikus. Tikus ICR betina dibahagikan secara rawak kepada 3 kumpulan (n= 8 tikus setiap kumpulan): Kawalan induksi dengan DMBA/TPA, kawalan negatif (aseton) dan kumpulan pra-rawatan oleuropein. Bagi kumpulan rawatan, tikus akan diinduksi dengan DMBA (200 nmol) diikuti dengan pra-rawatan oleuropein (10 mg/kg) sebelum diaruh dengan TPA (20 nmol) secara topikal atas bahagian dorsal sehingga 10 minggu. Analisis histopatologi menunjukkan bahawa kumpulan yang dirawat dengan oleuropein kelihatan kekurangan ketebalan epidermis hiperplasia, justeru, menghentikan peringkat galakan tumor berbanding dengan ketebalan hiperplasia dan ketidakaturan susunan epidermis pada kumpulan kawalan DMBA/TPA. Data juga menunjukkan bahawa kumpulan pra-rawatan oleuropein menghasilkan kenaikan signifikan kadar apoptosis (p<0.05) melalui pengaktifan caspase-3 bertanda berbanding kumpulan DMBA/TPA. Menariknya, tahap MDA menunjukkan pengurangan signifikan (p<0.05) dalam kumpulan pra-rawatan oleuropein berbanding kumpulan DMBA/TPA. Seterusnya, pra-rawatan oleuropein menyebabkan penurunan signifikan pada paras GSH (p<0.05) beserta peningkatan signifikan pada paras SOD (p<0.05) berbanding kumpulan DMBA/TPA. Secara keseluruhannya, kajian ini menunjukkan bahawa oleuropein berpotensi bertindak sebagai agen kemohalang melalui aktiviti apoptotik dan pertahanan antioksidan pada peringkat promosi tumor dalam karsinogenesis kulit.

Kata kunci: Kanser kulit; karsinogenesis; kemohalang; oleuropein; promosi tumor

RUJUKAN

Abel, E.L., Angel, J.M., Kiguchi, K. & DiGiovanni, J. 2009. Multi-stage chemical carcinogenesis in mouse skin: Fundamentals and applications. Nature Protocols 4(9): 1350-1362.

Anter, J., Fernandez-Bedmar, Z., Villatoro-Pulido, M., Demyda- Peyras, S., Moreno-Millan, M., Alonso-Moraga, A., Munoz- Serrano, A. & Luque de Castro, M.D. 2011. A pilot study on the DNA-protective, cytotoxic and apoptosis-inducing properties of olive-leaf extracts. Mutation Research 723(2): 165-170.

Barbaro, B., Toietta, G., Maggio, R., Arciello, M., Tarocchi, M., Galli, A. & Balsano, C. 2014. Effects of the olive-derived polyphenol oleuropein on human health. International Journal of Molecular Sciences 15(10): 18508-18524.

Basri, D.F., Alamin, Z.A.Z. & Chan, K.M. 2016. Assessment of cytotoxicity and genotoxicity of stem bark extracts from Canarium odontophyllum Miq. (dabai) against HCT 116 human colorectal cancer cell line. BMC Complementary and Alternative Medicine 16(36): 1-12.

Beyer, W.F. & Fridovich, I. 1987. Assaying for superoxide dismutase activity: Some large consequences of minor changes in condition. Analytical Biochemistry 161: 559-566.

Bishayee, A. & Sethi, G. 2016. Bioactive natural products in cancer prevention and therapy: Progress and promise. Seminars in Cancer Biology 40-41: 1-3.

DelBoccio, P., DiDeo, A., DeCurtis, A., Celli, N., Lacoviello, L. & Rotilio, D. 2003. Liquid chromatography-tandem mass spectrometry analysis of oleuropein and its metabolite hydroxytyrosol in rat plasma and urine after oral administration. Journal of chromatography B, Analytical Technologies in the Biomedical and Life Sciences 785(1): 47-56.

Elamin, M.H., Daghestani, M.H., Omer, S.A., Elobeid, M.A., Virk, P., Al-Olayan, E.M., Hassan, Z.K., Mohammed, O.B. & Aboussekhra, A. 2013. Olive oil oleuropein has anti-breast cancer properties with higher efficiency on ER-negative cells. Food and Chemical Toxicology 53: 310-316.

Ellman, G.L. 1959. Tissue sulfhydryl groups. Archives of Biochemistry and Biophysics 82: 70-77.

Friesen, C. & Debatin, K.M. 2004. A critical role of glutathione in determining apoptosis sensitivity and resistance in leukemia cells. Cell Death and Differentiation 1: s73-s85.

Harun, Z. & Ghazali, A.R. 2012. Potential chemoprevention activity of pterostilbene by enhancing the detoxifying enzymes in the HT-29 cell line. Asian Pacific Journal of Cancer Prevention 13(12): 6403-6407.

Ighodaro, O.M. & Akinloye, O.A. 2017. First line defence antioxidants-superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX): Their fundamental role in the entire antioxidant defence grid. Alexandria Journal of Medicine doi: 10.1016/j.ajme.2017.09.001

Kim, M.O., Kim, S.H., Shin, M.J., Yu, D.H., Kim, B.S., Chang, K.T., Lee, S., Park, Y.B., Lee, T.H. & Rhoo, Z.Y. 2007. DMBA/TPA-induced tumour formation is aggravated in human papillomavirus type 16 E6/E7 transgenic mouse skin. Oncology Research 16(7): 325-332.

Kimura, Y. & Sumiyoshi, M. 2009. Olive leaf extract and its main component oleuropein prevent chronic ultraviolet B radiation-induced skin damage and carcinogenesis in hairless mice. Journal of Nutrition 139(11): 2079-2086.

Konoshima, T. & Takasaki, M. 2000. Anti-tumor-promoting activities (cancer chemopreventive activities) of natural products. Studies in Natural Products Chemistry 24: 215-267.

Lagman, M., Ly, J., Saing, T., Singh, M.K., Tudela, E.V., Morris, D., Chi, P.T., Ochoa, C., Sathananthan, A. & Venketaraman, V. 2015. Investigating the causes for decreased levels of glutathione in individuals with type II diabetes. Plos ONE 10(3): e0118436.

Lindner, D. 2014. Animal models and the tumor microenvironments: Studies of tumor-host symbiosis. Seminars in Oncology 41(2): 146-155.

Liu, Y., Borchert, G.L. & Donals, S.P. 2005. MnSOD inhibits proline oxidase-induced apoptosis in colorectal cancer cells. Carcinogenesis 26: 1335-1342.

Masre, S.F., Rath, N., Olson, M.F. & Greenhalgh, D.A. 2017. ROCK2/rasHa co-operation induces malignant conversion via p53 loss, elevated NF-ĸβ and tenascin C-associated rigidity, but p21 inhibits ROCK2/NF- ĸβ-mediated progression. Oncogene 36: 2529-2542.

Micol, V., Caturla, N., Perez-Fons, L., Mas, V., Perez, L. & Estepa, A. 2005. The olive leaf extract exhibits antiviral activity against viral haemorrhagic septicaemia rhabdovirus (VHSV). Antiviral Research 66(2-3): 129-136.

Milanizadeh, S., Bigdeli, M.R., Rasoulian, B. & Amani, D. 2014. The effects of olive leaf extract on antioxidant enzymes activity and tumor growth in breast cancer. Thrita 3(1): e12914.

Neagu, M., Caruntu, C., Constantin, C., Boda, D., Zurac, S., Spandidos, D. & Abd Tsatsakis, A.M. 2016. Chemically induced skin carcinogenesis: Updates in experimental models (review). Oncology Reports 35(5): 2516-2528.

Neergheen, V.S., Bahorun, T., Taylor, E.W., Jen, L.S. & Aruoma, O.I. 2010. Targeting specific cell signalling transduction pathways by dietary and medicinal phytochemicals in cancer chemoprevention. Toxicology 278(2): 229-241.

Omar, S.H. 2010. Oleuropein in olive and its pharmacological effects. Scientia Pharmaceutica 78(2): 133-154.

Qabaha, K., AL-Rimawi, F., Qasem, A. & Naser, S.A. 2017. Oleuropein is responsible for the major anti-inflammatory effects of olive leaf extract. Journal of Medicinal Food doi: org/10.1089/jmf.2017.0070.

Rundhaug, J.E. & Fischer, S.M. 2010. Molecular mechanisms of mouse skin tumor promotion. Cancers 2: 436-482.

Salleh, S.A., Rajab, N.F., Abdullah, N.R., Ismail, Z., Mouatt, P., Dowell, A. & Muhamad, S. 2011. In vitro chemopreventive activity of an ethyl acetate fraction derived from hot water extract of Orthosiphon stamineus in HepG2 cells. Journal of Medicinal Plant Research 5(10): 1892-1899.

Shamshoum, H., Vlavcheski, F. & Tsiani, E. 2017. Anticancer effects of oleuropein. Biofactors 43(4): 517-528.

Stocks, J. & Dormandy, T.L. 1971. The autoxidation of human red cell lipids induced by hydrogen peroxide. British Journal of Haematology 20: 95-111.

Strzelczyk, J.K. & Wiczkowski, A. 2012. Oxidative damage and carcinogenesis. Contemporary Oncology 16(3): 230-233.

Thorsen, M., Jacobson, T., Vooijs, R., Navarrete, C., Bliek, T., Schat, H. & Tamas, M.J. 2012. Glutathione serves an extracellular defence function to decrease arsenite accumulation and toxicity in yeast. Molecular Microbiology 84(6): 1177-1188.

Yan, C.M., Chai, E.Q., Cai, H.Y., Miao, G.Y. & Ma, W. 2015. Oleuropein induces apoptosis via activation of caspases and suppression of phosphatidylinositol 3-kinase/protein kinase B pathway in HepG2 human hepatoma cell line. Molecular Medicine Reports 11(6): 4617-4624.

Yoshikawa, T. & Naito, Y. 2002. What is oxidative stress. Japan Medical Association Journal 45(7): 271-276.

Zaila, C.F.S., Zuraina, M.Y.F., Norfazlina, M.N., Mun, L.L., Nurshahirah, N., Florinsiah, L. & Rajab, N.F. 2013. Antiproliferative effect of Lignosus rhinocerotis, the Tiger Milk mushroom on HCT 116 human colorectal cancer cell. The Open Conference Proceeding Journal 4(2): 65-70.

*Pengarang untuk surat-menyurat; email: sitifathiah@ukm.edu.my