Sains Malaysiana 51(1)(2022): 217-237

http://doi.org/10.17576/jsm-2022-5101-18

Zingiber officinale and Piper betle Extracts Enhanced the Chemopreventive Effect against Colon Cancer Cells by Targeting Caspase-Mediated Apoptosis

  (Ekstrak Zingiber officinale dan Piper betle Meningkatkan Kesan Kemohalang terhadap Sel Kanser Kolon dengan Menyasarkan Apoptosis Diperantara-Kaspase)

YASMIN ANUM MOHD YUSOF¹, SHAILAH ABDULLAH², NUR FATIN NABILAH MOHD SAHARDI², WAN ZURINAH WAN NGAH² & SUZANA MAKPOL²*

¹Faculty of Medicine and Defense Health, Universiti Pertahanan Nasional Malaysia, Kem Sungai Besi, 57000 Kuala Lumpur, Federal Territory, Malaysia

²Department of Biochemistry, Level 17 Preclinical Building, Universiti Kebangsaan Malaysia Medical Center, Jalan Yaacob Latif, Bandar Tun Razak, 56000 Cheras, Kuala Lumpur, Federal Territory, Malaysia

Received: 22 April 2020/Accepted: 19 April 2021

ABSTRACT

Food regimens and herbs have been a target of scientific research in treating cancer. This study aimed to elucidate the effect of Zingiber officinale (ginger) and Piper betle (PB) alone or in combination on the colorectal cancer cell lines HCT116 and HT29. The chemopreventive effect of these extracts was determined by performing cell viability assay, cell cycle analysis, apoptosis analysis, and caspase 3 and 8 activity assays in HCT116 and p53-deficient HT29 cells. Ginger and PB extracts inhibited the proliferation of both cancer cell lines dose-dependently, but the combined extracts inhibited the growth of cancer cells synergistically with an IC50 PB plus ginger < IC50 PB < IC50 ginger. Similarly, the effect of the combined extracts on apoptosis of both cancer cell lines was higher than that of each individual extract. Consistent with the enhancement of caspase activity, both extracts increased the expression of BAX protein, while Bcl-2 protein was decreased. Flow cytometry analysis indicated that the combined extracts arrested a higher percentage of colon cancer cells at the G0/G1 phase with a concomitant decrease in cells in the S phase, indicating that the combined extracts inhibited human colon cancer cell growth by cell cycle arrest at the G0/G1 phase. The inhibitory effect of these combined extracts on cellular proliferation and the induction of apoptosis was synergistic, showing a significant combination index (CI) value < 1.0. The combined extracts of ginger and PB may serve as a potential chemopreventive agent against colon cancer through the induction of caspase-mediated apoptosis.

Keywords: Apoptosis; cell cycle; colorectal cancer; Piper betle; Zingiber officinale

ABSTRAK

Industri makanan dan herba telah menjadi salah satu tarikan dalam kajian saintifik untuk merawat kanser. Objektif kajian ini dijalankan adalah untuk menentukan kesan Zingiber officinale (halia) dan Piper betle (PB) secara individu atau gabungan terhadap titisan sel kanser kolorektum, HCT116 dan HT29. Kesan kemohalang kedua-dua ekstrak ini ditentukan melalui asai kebolehidupan, analisis kitaran sel, analisis apoptosis, asai aktiviti kaspase 3 dan 8 terhadap sel HCT116 dan sel HT29 yang kekurangan-p53. Ekstrak halia dan PB telah menghalang proliferasi untuk kedua-dua jenis sel kanser mengikut kepekatan dos, akan tetapi gabungan kedua-dua ekstrak telah menghalang pertumbuhan sel secara sinergistik dengan IC₅₀ gabungan PB dan halia < IC₅₀ PB < IC₅₀ halia. Kesan apoptosis terhadap sel kanser bagi rawatan gabungan kedua-dua ekstrak juga lebih tinggi berbanding rawatan secara individu. Selari dengan peningkatan aktiviti kaspase, kedua-dua ekstrak telah meningkatkan ekspresi protein BAX dan menurunkan pengekspresan protein Bcl-2. Analisis aliran sitometri menunjukkan bahawa rawatan gabungan kedua-dua ekstrak telah menyebabkan peningkatan peratusan yang tinggi terhadap kitaran sel kanser kolon pada fasa G₀/G₁ dan pada masa yang sama menyebabkan penurunan peratusan sel pada fasa S, membuktikan bahawa gabungan kedua-dua ekstrak telah menghalang pertumbuhan sel kanser pada fasa G₀/G_1. Kesan perencatan gabungan kedua-dua ekstrak terhadap proliferasi sel dan induksi apoptosis adalah secara sinergistik dengan nilai indeks gabungan (CI) < 1.0. Oleh yang demikian, gabungan ekstrak halia dan PB mungkin mempunyai potensi sebagai agen kemohalang bagi melawan kanser kolon dengan cara menyasarkan apoptosis-diperantara kaspase.

Kata kunci: Apoptosis; kanser kolorektal; kitaran sel; Piper betle; Zingiber officinale

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*Corresponding author; email: suzanamakpol@ppukm.ukm.edu.my