Antiproliferative activity and caspase enhancement properties of Annona muricata leaves extract against colorectal cancer cells

Lili Indrawati; Purwantyastuti Ascobat; Budiman Bela; Murdani Abdullah; Ingrid S. Surono; Suwijiyo Pramono

doi:10.13181/mji.v25i3.1449

Authors

Lili Indrawati Department of Pharmacology, Faculty of Medicine, Christian University of Indonesia, Jakarta
Purwantyastuti Ascobat Department of Pharmacology, Faculty of Medicine, Universitas Indonesia, Jakarta
Budiman Bela Department of Microbiology, Faculty of Medicine, Universitas Indonesia, Jakarta
Murdani Abdullah Department of Internal Medicine, Faculty of Medicine, Universitas Indonesia, Cipto Mangunkusumo Hospital, Jakarta
Ingrid S. Surono Department of Food Technology, Faculty of Engineering, Bina Nusantara University, Jakarta
Suwijiyo Pramono Department of Pharmaceutical Biology, Faculty of Pharmacy, Gadjah Mada University, Yogyakarta

DOI:

https://doi.org/10.13181/mji.v25i3.1449

Keywords:

A. muricata leaves, cytotoxicity, colorectal cancer cell line, ex vivo, in vitro

Abstract

Background: The prevalence of colorectal cancer is rising in Asia including Indonesia. Annona muricata tea leaves, that is traditionally used for maintaining health, and lately being used by cancer patients. The objectives of this study is to investigate its effects in human colorectal cancer cell in vitro and ex vivo.

Methods: Thirty patients with colorectal cancer (CRC) were enrolled in a randomized double-blind placebo-controlled trial. They were equally divided into two groups: those treated with 300 mg A. muricata leaf extract and placebo daily for 8 weeks. Serum from supplemented CRC patients of both groups was compared for caspase 9 and caspase 8 enhancement activity. Antiproliferative effect of water extract of A. muricata leaves and its fractions were evaluated against colorectal cancer cell line (DLD-1 and COLO 205) compared with 5-fluorouracil and placebo, the dose range was 62.5-2,000 μg/mL. Method used was 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Data were analyzed by Mann-Whitney U test. The p value was set at 0.05.

Results: Ethanol-soluble fraction of A. muricata leaves extract water extract (ESFAM) leaves extract had cytotoxicity effects on DLD-1 as well as COLO 205 cell line, as shown by the lower IC 50 compared to 5-fluorouracil and placebo, 20.59 μg/mL and 654.9μg/mL, respectively. Serum of subjects supplemented with extract significantly induced caspase 9 (p=0.001) activity of DLD-1 colorectal cancer cell line, but not for caspase 8 activity (p=0.372).

Conclusion: The study's results suggest the cytotoxicity potential of A. muricata leaves extract in in vitro and ex vivo studies.

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References

Yee YK, Tan VP, Chan P, Hung IF, Pang R, Wong BC. Epidemiology of colorectal cancer in Asia. J Gastroenterol Hepatol. 2009;24(12):1810-6. http://dx.doi.org/10.1111/j.1440-1746.2009.06138.x

Pourhoseingholi MA. Increased burden of colorectal cancer in Asia. World J Gastrointest Oncol. 2012;4(4):68-70. http://dx.doi.org/10.4251/wjgo.v4.i4.68

Soeripto, Indrawati, Indrayani. Gastro-intestinal cancer in Indonesia. Asian Pac J Cancer Prev. 2003;4(4):289-96.

Psahoulia FH, Drosopoulos KG, Doubravska L, Andera L, Pintzas A. Quercetin enhances TRAIL-mediated apoptosis in colon cancer cells by inducing the accumulation of death receptors in lipid rafts. Mol Cancer Ther. 2007;6(9):2591-9. http://dx.doi.org/10.1158/1535-7163.MCT-07-0001

Khan AQ, Bury JP, Brown SR, Riley SA, Corfe BM. Keratin 8 expression in colon cancer associates with low faecal butyrate levels. BMC gastroenterol. 2011;11(1):2-10. http://dx.doi.org/10.1186/1471-230X-11-2

Yu DQ. Recent works on anti-tumor constituent from Annonaceae plants in China. Pure Appl Chem.1999;71(6):1119-22. http://dx.doi.org/10.1351/pac199971061119

Champy P, Melot A, Guérineau Eng V,, Gleye C, Fall D, Höglinger GU, et al. Quantification of acetogenins in Annona muricata linked to atypical parkinsonism in guadeloupe. Mov Disord. 2005;20(12):1629-33. http://dx.doi.org/10.1002/mds.20632

Zuhud E. Kanker Lenyap Berkat Sirsak. 1st ed. Indah Y, editor. Jakarta: PT AgroMedia Pustaka; 2011. p. 37-39. Indonesian.

García-Aguirre KK, Zepeda-Vallejo LG, Ramón-Gallegos E, Alvárez-González I, Madrigal-Bujaidar E. Genotoxic and cytotoxic effects produced by acetogenins obtained from Annona cherimolia Mill. Biol Pharm Bull. 2008;31(12):2346-9. http://dx.doi.org/10.1248/bpb.31.2346

Gupta A, Pandey S, Shah D, Yadav J, Seth N. Annonaceous acetogenins: the unrevealed area for cytotoxic and pesticidal activities. Syst Rev Pharm. 2011;2(2):104-9. http://dx.doi.org/10.4103/0975-8453.86299

Taylor L. Technical data report for graviola Annona muricata. In: Taylor L, editor. Herbal Secretes of the Rainforest. Austin: Rain Tree; 2002. p. 1-43.

Marino DC, Sabino LZ, Armando J Jr, Ruggiero Ade A, Moya HD. Analysis of the polyphenols content in medicinal plants based on the reduction of Cu(II)/bicinchoninic complexes. J Agric Food Chem. 2009;57(23):11061-6. http://dx.doi.org/10.1021/jf902197p

Adewole SO, Ojewole JA. Protective effects of Annona muricata Linn. (Annonaceae) leaf aqueous extract on serum lipid profiles and oxidative stress in hepatocytes of streptozotocin-treated diabetic rats. Afr J Tradit Complement Altern Med. 2008;6(1):30-41.

Ezirim AU, Okachi VI, James AB, Adebeshi OA, Ogunnowo S, Odeghe O. Induction of apoptosis in myelogenous leukemic K562 cells by ethanolic leaf extract of Annona muricata. Global J Res Med Plants & Indigen Med. 2013;2(3):142-51.

Alley MC, Scudiero DA, Monks A, Hursey ML, Czerwinski MJ, Fine DL, et al. Feasibility of drug screening with panels of human tumor cell lines using a microculture tetrazolium assay. Cancer Res. 1988;48(3):589-601.

Vistica DT, Skehan P, Scudiero D, Monks A, Pittman A, Boyd MR. Tetrazolium-based assays for cellular viability: a critical examination of selected parameters affecting formazan production. Cancer Res. 1991;51(10):2515-20.

Schmid J, Dussmann H, Boukes GJ, Flanagan L, Lindner AU, O'Connor CL, et al. Systems analysis of cancer cell heterogeneity in caspase-dependent apoptosis subsequent to mitochondrial outer membrane permeabilization. J Biol Chem. 2012;287(49):41546-59. http://dx.doi.org/10.1074/jbc.M112.411827

Kim GS, Zeng L, Alali F, Rogers LL, Wu FE, McLaughlin JL, et al. Two new mono-tetrahydrofuran ring acetogenins, annomuricin E and muricapentocin, from the leaves of Annona muricata. J Nat Prod. 1998;61(4):432-6. http://dx.doi.org/10.1021/np970534m

Chang FR, Liaw CC, Lin CY, Chou CJ, Chiu HF, Wu YC. New adjacent Bis-tetrahydrofuran Annonaceous acetogenins from Annona muricata. Planta Med. 2003;69(3):241-6. http://dx.doi.org/10.1055/s-2003-38485

De Angelis PM, Svendsrud DH, Kravik KL, Stokke T. Cellular response to 5-fluorouracil (5-FU) in 5-FU-resistant colon cancer cell lines during treatment and recovery. Mol Cancer. 2006;5:20-44. http://dx.doi.org/10.1186/1476-4598-5-20

Bartke T, Siegmund D, Peters N, Reichwein M, Henkler F, Scheurich P, et al. p53 upregulates cFLIP, inhibits transcription of NF-kappaB-regulated genes and induces caspase-8-independent cell death in DLD-1 cells. Oncogene. 2001;20(5):571-80. http://dx.doi.org/10.1038/sj.onc.1204124

Dodge MW. Synthesis of annonaceous acetogenins via asymmetric double cycloetherification [dissertation]. United States: University of Wisconsin-Madison; 2008.

Xu ZF, Wei XY, Xie HH, Yang RZ. Inhibitory activities of three annonaceous acetogenins on NADH oxidase of chicken liver mitochondria. Biol Pharm Bull. 2003;26(5):729-32. http://dx.doi.org/10.1248/bpb.26.729