Sains Malaysiana 52(10)(2023): 2785-2801

http://doi.org/10.17576/jsm-2023-5210-05

The Phytochemical Components of Kelantan Grown Annona muricata Leaves and Its Anti-Proliferative Properties on MCF-7 Breast Cancer Cells

(Komponen Fitokimia Daun Annona muricata Ditanam di Kelantan dan Sifat Anti-Proliferatifnya terhadap Sel Kanser Payudara MCF-7)

NORZILA ISMAIL¹, HARISHINI RAJARATINAM², KHAIRUNNISA CHE GHAZALI³, SITI FARHANAH MOHD SALLEH¹, WAN SURIYANI FALIQ ADEEBA WAN IBRAHIM⁴, MAYA MAZUWIN YAHYA³ & WAN ZAINIRA WAN ZAIN³

¹Department of Pharmacology, School of Medical Sciences, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, Malaysia

²School of Health Sciences, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, Malaysia

³Department of Surgery, Hospital Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, Malaysia

⁴Faculty of Bioengineering & Technology, Universiti Malaysia Kelantan, Jeli Campus, 17600 Jeli Kelantan, Malaysia

Received: 13 June 2022/Accepted: 26 September 2023

Abstract

Annona muricata leaves exhibit anti-cancer properties against breast cancer. In this study, the phytochemical components, anti-proliferative activity, and cytotoxicity of Kelantan (Malaysia) grown Annona muricata leaves were evaluated using MCF-7 breast cancer cell line. The preparation of Annona muricata leaves extracts was performed with various organic solvents (ethyl acetate, n-hexane, methanol, and aqueous) using Soxhlet method. Each extract was analysed using gas chromatography-mass spectrometry (GCMS) for phytochemical analysis and characterised with Wiley and NIST library searchers. The effects of all extracts on cell proliferation were analysed using MTT assay. The anti-proliferative activity was determined by evaluating the IC₅₀ value. Based on the lowest IC₅₀value, further experiments were done to observe the apoptosis effect using the Annexin V-FITC technique. While for cell cycle analysis, Cycletest Plus DNA Reagent Kit was used. Both were further determined using flow cytometry analysis. The ethyl acetate-based Annona muricata extract contained terpenoids, phenolic compounds, and alkaloids. Ethyl acetate extract of Annona muricata exhibited a significant cytotoxic effect on MCF-7 breast cancer cells. The most potent IC₅₀ value obtained was 21.8 ± 3.85 µg/mL. The changes in morphology were more profound after 72 h of treatment with ethyl acetate extract of Annona muricata, including cell shrinkage. Flow cytometry analysis showed that it was induced in early and late apoptosis in vitro and arrest at G₀/G₁ cell cycle phase in time-dependant manners. In short, ethyl acetate extract of Annona muricata exhibited a range of phytochemical components, anti-proliferative activity and cytotoxic effect on MCF-7 breast cancer cells.

Keywords: Annona muricata; apoptosis; breast cancer; cytotoxicity; MCF-7

Abstrak

Daun Annona muricata menunjukkan sifat anti-kanser terhadap kanser payudara. Dalam kajian ini, komponen fitokimia, aktiviti anti-proliferatif dan kesitotoksikan daun Annona muricata yang ditanam di Kelantan (Malaysia) dinilai menggunakan titisan sel kanser payudara MCF-7. Penyediaan ekstrak daun Annona muricata dilakukan dengan pelbagai pelarut organik (etil asetat, n-heksana, metanol dan akueus) dengan menggunakan kaedah Soxhlet. Setiap ekstrak dianalisis menggunakan spektrometri jisim kromatografi gas (GCMS) untuk analisis fitokimia dan dicirikan dengan pencari perpustakaan Wiley dan NIST. Kesan semua ekstrak pada pertumbuhan sel telah dianalisis menggunakan asai MTT. Aktiviti anti-proliferatif ditentukan melalui nilai IC₅₀. Berdasarkan nilai IC₅₀ yang paling rendah, uji kaji lanjutan telah dilakukan untuk memerhatikan kesan apoptosis menggunakan teknik Annexin V-FITC. Manakala untuk analisis kitaran sel, Cycletest Plus DNA Reagent Kit digunakan. Keputusan daripada kedua-dua uji kaji dikukuhkan dengan kaedah analisis sitometri aliran. Ekstrak Annona muricata berasaskan etil asetat mengandungi terpenoid, sebatian fenol dan alkaloid. Ekstrak etil asetat Annona muricata menunjukkan kesan sitotoksik yang ketara pada sel kanser payudara MCF-7. Nilai IC₅₀ yang paling berkesan diperoleh ialah 21.8 ± 3.85 µg/mL. Perubahan morfologi sel lebih ketara selepas 72 jam rawatan dengan ekstrak etil asetat Annona muricata termasuk pengecutan sel. Analisis sitometri aliran menunjukkan bahawa ia telah diinduksi pada apoptosis awal dan lewat secara in vitro dan perencatan pada fasa kitaran sel G₀/G₁ secara berkadar langsung dengan masa. Ringkasnya, ekstrak etil asetat Annona muricata menunjukkan pelbagai komponen fitokimia, aktiviti anti-proliferatif dan kesan sitotoksik terhadap sel kanser payudara MCF-7.

Kata kunci: Annona muricata; apoptosis; kanser payudara; kesitotoksian; MCF-7

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*Corresponding author; email: zainira@usm.my