The effect of ethanolic leaves extract of soursop (Annona muricata L.) on human colorectal cancer cell line: cell viability and in silico study to cyclin D1 protein
Article Sidebar
Vol. 10 No. 2 (2019)
November 15, 2019
December 31, 2019
Abstract
Latar Belakang: Kanker kolorektal merupakan transformasi patologis dari epitel kolon dan rektum normal menjadi massa jaringan abnormal, perubahan ini terjadi karena ekspresi berlebih dari protein cyclin D1 yang menginduksi proliferasi sel kolorektal secara berlebihan. Pengobatan dan pencegahan kanker kolorektal dapat dilakukan secara alami dengan mengonsumsi ekstrak daun Annona muricata L. (sirsak). Sirsak dikenal karena banyak komponen fitokimia yang berfungsi sebagai anti kanker.
Metode: Penelitian ini menggunakan sel kanker kolorektal HT-29 yang diberi ekstrak etanol daun sirsak dan 5 Fluorourasil (5-FU). Tujuannya untuk menemukan konsentrasi sitotoksisitas yang dapat menghambat 50% populasi sel HT-29 (CC50) dan konsentrasi yang didapat sebelumnya akan diuji dengan metode uji MTT. Analisis docking molekuler dilakukan antara molekul-molekul dari ekstrak etanol daun sirsak terhadap protein Cyclin D1 menggunakan perangkat lunak molecular operating environment (MOE) 2013.08.
Hasil: CC50 ekstrak etanol daun sirsak adalah 278 μg / mL dan 5-FU adalah 88 μg / mL. Persentase terendah sel HT-29 yang layak adalah 2 x CC50 setelah perlakuan ekstrak etanol daun sirsak (40,4 ± 1,3%) dibandingkan dengan 5-FU (52,8 ± 4,3%), kontrol pelarut ( 97,2 ± 1,4%), dan kontrol sel (100%). Analisis docking molekuler untuk protein cyclin D1 diperoleh asam N-hexadecanoic dan molekul phytol sebagai kandidat yang baik untuk menghambat protein cyclin D1.
Kesimpulan: Ekstrak etanol daun sirsak dapat menurunkan viabilitas sel kultur kanker kolon HT-29 dan berdasarkan analisis molekular docking dilihat dari energi bebas gibbs (ΔG) dan afinitas tertinggi (pKi) diperoleh N-hexadecanoic dan molekul phytol sebagai penghambat protein cyclin D1. (Health Science Journal of Indonesia 2019;10(2):96-102)
Kata Kunci: Kanker kolorektal HT-29, ekstrak etanol daun sirsak, viabilitas sel, molecular docking, cyclin D1
Abstract
Introduction: Colorectal cancer is a pathological transformation of normal colon and rectum epithelial that becomes an abnormal tissue mass, due to the overexpression of cyclin D1 protein that inducing excessive proliferation of colorectal cell. The treatment and prevention of colorectal cancer could be done naturally by consuming leaves extract of Annona muricata L. (soursop). Soursop is known for many phytochemical components that serve as an anti-cancer.
Methods: This study was used HT-29 colorectal cancer cell that treated with ethanolic leaves extract of soursop and 5-Fluorourasil (5-FU) to find the cytotoxicity concentration that can inhibit 50% of HT-29 cell population (CC50) and the next concentrations of them were treated for next treatment with MTT assay. Molecular docking analysis of the compounds of ethanolic leaves extract of soursop to cyclin D1 protein used molecular operating environment (MOE) 2013.08 software.
Results: CC50 of ethanolic leaves extracts of soursop was 278 μg/mL dan 5-FU was 88 μg/mL. The lowest percentage of viable HT-29 cell was 2 x CC50 after ethanolic leaves extract of soursop treatment (40,4±1,3%) was compared to 5-FU (52,8±4,3%), solvent control (97,2±1,4%), and cells control (100%). Analysis of molecular docking to cyclin D1 protein was obtained N-hexadecanoic acid and phytol molecules as good candidates to inhibit cyclin D1 protein.
Conclusions: The ethanolic leaves extract of soursop could be a good alternative treatment for colorectal cancer and its compounds had ability to inhibit cyclin D1 protein (the highest gibbs free energy (ΔG) and affinity (pKi)). (Health Science Journal of Indonesia 2019;10(2):96-102)
Keywords: Colorectal cancer, ethanolic leaves extract of soursop, cell viability, molecular docking, cyclin D1
References
Kuipers EJ, Grady WM, Lieberman D, Seufferlein T, Sung JJ, Boelens PG, et al. Colorectal cancer. Nat Rev Dis Primers. 2015 Nov 5;1:15065.
McGuire S. World Cancer Report 2014. Geneva, Switzerland: World Health Organization, International Agency for Research on Cancer, WHO Press, 2015. Adv Nutr. 2016;7(2):418-9.
Libutti SK, Salz LB, Willett CG, Levine RA. Chapter 57: Cancer of the colon. In: DeVita VT, Lawrence TS, Rosenberg SA, eds. DeVita, Hellman, and Rosenberg’s Cancer: Principles and Practice of Oncology. 10th ed. Philadelphia, Pa: Lippincott Williams & Wilkins; 2015.
George VC, Kumar DN, Rajkumar V, Suresh P, Ashok R. Quantitative assessment of the relative antineoplastic potential of then-butanolic leaf extract of Annona Muricata Linn. in normal and immortalized human cell lines. Asian Pacific J Can Prev. 2012;13:699-704.
Moghadamtousi SZ, Karimian H, Rouhollahi E, Paydar MJ, Fadaeinasab M, Kadir HA. Annona muricata leaves induce G1 cell cycle arrest and apoptosis through mitochondria-mediated pathway in human HCT-116 and HT-29 colon cancer cells. J Ethnopharm. 2014;156:277-89.
Gajalakshmi S, Vijayalakshmi S, Rajeshwari Devi V. Phytochemical and pharmacological properties of annona muricata: a review. Int J Pharm Sci. 2012;4(2):3-6.
Patel S, Patel JK. A review on a miracle fruits of annona muricata. J Pharmacog Phytochem. 2016;5(1):137-48.
Abdullah M, Syam AF, Meilany S, Laksono B, Prabu OG, Bekti HR, et al. The value of caspase-3 after the application of annona muricata leaf extract in COLO-205 colorectal cancer cell line. Gastroent Res Pract. 2017;1-5.
Indrawati L, Pramono S, Ascobat P, Bela B, Abdullah M, Surono I. Cytotoxic activity of soursop “Annona muricata” leaves extracts and their phytochemical contents. JGPT. 2017;02(9):35-40.
Ko YM, Wu TY, Wu YC, Chang FR, Guh JY, Chuang LY. Annonacin induces cell cycle-dependent growth arrest and apoptosis in estrogen receptor-α-related pathways in MCF-7 cells. J Ethnopharm. 2011;137:1283–290.
John RR, Malathi N, Ravindran C, Anandan S. Mini review: multifaceted role played by cyclin D1 in tumor behavior. Indian J Dent Res 2017;28:187-92.
Li Y, Wei J, Xu C, Zhao Z, You T. Prognostic significance of cyclin D1 expression in colorectal cancer: a meta analysis of observational studies. PLoS One. 2014;9(4):e94508.
Nie J, Liu L, Zheng W, Chen L, Wu X, Xu Y, et al. MicroRNA-365, down-regulated in colon cancer, inhibits cell cycle progression and promotes apoptosis of colon cancer cells by probably targeting Cyclin D1 and Bcl-2. Carcinogenesis. Jan 2012;33(1):220-25.
Wangefjord S, Manjer J, Gaber A, Nodin B, Eberhard J, Jirström K. Cyclin D1 expression in colorectal cancer is a favorable prognostic factor in men but not in women in a prospective, population-based cohort study. Biol Sex Differ. 2011;2:10.
Sameer AS, Parray FQ, Dar MA, Nissar S, Banday MZ, Rasool S, et al. Cyclin D1 G870A polymorphism and risk of colorectal cancer: a case control study. Mol Med Rep. 2013;7(3):811-5.
Bahuguna A, Khan I, Bajpai VK, Kang SC. MTT Assay to evaluate the cytotoxic potential of a drug. Bangladesh J Pharmacol. 2017;12:115-8.
Gavamukulya Y, Abou-Elella F, Wamunyokoli F, El-Shemy HA.GC-MS analysis of bioactive phytochemicals present in ethanolic extracts of leaves of Annona muricata: a further evidence for its medicinal diversity. Pharmacog J. Sep 2015;7(5):300-4.
Moghadamtousi SZ, Rouhollahi E, Karimian H, Fadaeinasab M, Firoozinia M, Abdullah MA, et al. The chemopotential effect of annona muricata leaves against azoxymethane-induced colonic aberrant crypt foci in rats and the apoptotic effect of acetogenin annomuricin E in HT-29 cells: a bioassay-guided approach. PLoS ONE 2015;10(4): e0122288.
Wijayanti PA, Puspaningtyas AR, Pangaribowo DA. Uji sitotoksisitas dan proliferasi senyawa 1-(4- Trifluorometilbenzoiloksimetil)-5-Fluorourasil terhadap sel kanker payudara MCF-7. e-Jurnal Pustaka Kesehatan. September 2015;3(3):1-5. Indonesian.
Kim YS, Hong J, Sym SJ, et al. Oxaliplatin, 5-fluorouracil and leucovorin (FOLFOX-4) combination chemotherapy as a salvage treatment in advanced gastric cancer. Cancer Res Treat. 2010;42(1):24–9.
Calabro-Jones PM, Byfield JE, Ward JF, Sharp TR. Time-dose relationships for 5-fluorouracil cytotoxicity against human epithelial cancer cells in vitro. Cancer Res. 1982 Nov;42(11):4413-20.
De Angelis PM, Svendsrud DH, Kravik KL, Stokke T. Cellular response to 5-fluorouracil (5-FU) in 5-FUresistant colon cancer cell lines during treatment and recovery. Mol Canc. May 2006;5(20):1-25.
Noordhuis P, Holwerda U, Van der Wilt CL, Van Groeningen CJ, Smid K, Meijer S, et al. 5-Fluorouracil incorporation into RNA and DNA in relation to thymidylate synthase inhibition of human colorectal cancers. Annals of Oncology. 2004;15:1025–32.
Lee HS, Jo S, Lim HS, Im W. Application of binding free energy calculations to prediction of binding modes and affinities of MDM2 and MDMX inhibitors. J Chem Inf Model. 2012;52(7):1821–32.
National Center for Biotechnology Information. PubChem Database. G1/S-specific cyclin-D1 [Homo sapiens], https://www.ncbi.nlm.nih.gov/protein/NP_444284.1 (accessed on May 10, 2019).
Alao JP. The regulation of cyclin D1 degradation: roles in cancer development and the potential for therapeutic invention. Mol Cancer. 2007;6:24.
Lee HS, Jo S, Lim HS, Im W. Application of binding free energy calculations to prediction of binding modes and affinities of MDM2 and MDMX inhibitors. J Chem Inf Model. 2012;52(7):1821-32.
Ramírez D, Caballero J. Is it reliable to take the molecular docking top scoring position as the best solution without considering available structural data?. Molecules. 2018;23(5):1038.
