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Sains
Malaysiana 53(9)(2024): 3159-3171
http://doi.org/10.17576/jsm-2024-5309-20
Mechanism of Damnacanthal
Induced Apoptosis in CEM-SS Cell Line
(Mekanisme Apoptosis
Teraruh Damnacanthal dalam Titisan Sel CEM-SS)
BANULATA
GOPALSAMY, SAIFUL YAZAN LATIFAH* & HISYAM ABDUL HAMID
Department
of Biomedical Science, Faculty of Medicine and Health Sciences, Universiti
Putra Malaysia, 43400 UPM Serdang,
Selangor, Malaysia
Received: 9 January 2024/Accepted: 5 August 2024
Leukaemia,
is cancer of organs that is responsible to produce blood specifically the
lymphatic system and bone marrow. Due to the harsh effects of currently used
cancer drugs, damnacanthal, an anthraquinone obtained from the roots of Morinda
elliptica is tested as a potential anticancer agent. This study reports on
the participation of the p53, Bcl-2 and Bax in the apoptosis induced by of
damnacanthal, on T-lymphoblastic leukaemia (CEM-SS) cell. Cell viability and
morphology was tested with trypan blue assay, flow cytometry analysis detected
the apoptotic activity of damnacanthal, caspase colorimetric protease assay
tested the Caspase 2, 3, 6, 8, and 9’s involvement and Enzyme-linked
Immunosorbent Assay (ELISA) was carried out to quantify the Human p53, Bcl-2,
and Bax expression levels. Damnacanthal exhibited cytotoxicity at doses 10 and
30 µg/mL after 72 h of incubation. This study reports that damnacanthal
arrested the cell at G2/M phase and initiates the apoptotic activity in the cells
treated with 30 µg/mL of damnacanthal for 72 h through caspase 2 and 6
activation and not caspases 3, 8, and 9. Furthermore, this anthraquinone
induces apoptosis via p53-independent pathway. Damnacanthal also lowered Bcl-2
and increased Bax activity in CEM-SS cell lines. These anticancer properties of
damnacanthal makes it a potential agent to treat T-lymphoblastic leukaemia.
Keywords: Anticancer;
apoptosis; CEM-SS; damnacanthal
Abstrak
Leukemia adalah kanser bagi organ yang bertanggung jawab
menghasilkan darah, terutamanya sistem limfa dan sum sum tulang. Disebabkan
kesan yang buruk oleh ubat kanser yang sedia ada, damnacanthal, salah satu
antrakuinon yang diperoleh daripada akar Morinda elliptica telah diuji
sebagai agen anti kanser yang berpotensi. Penyelidikan ini melaporkan
penglibatan p53, Bcl-2 dan Bax dalam apoptosis aruhan
damnacanthal, ke atas sel T-limfoblastik leukemia (CEM-SS). Kemandirian sel dan
morfologi telah diuji dengan ujian tripan biru, analisis sitometri aliran
mengesan aktiviti apoptosis damnacanthal, ujian protease kolorimetrik caspase
menguji penglibatan Caspase 2, 3, 6, 8 dan 9 dan Ujian Imunosorben Berkaitan
Enzim (ELISA) telah dijalankan untuk mengukur tahap pengekspresan p53, Bcl-2
dan Bax manusia. Damnacanthal menunjukkan sitotoksisiti pada dos 10 dan 30
µg/mL selepas 72 jam pengeraman. Kajian ini melaporkan bahawa damnacanthal
menahan sel pada fasa G2/M dan memulakan aktiviti apoptosis dalam sel yang
dirawat dengan 30 µg/mL damnacanthal selama 72 jam melalui pengaktifan caspase
2 dan 6 dan bukan caspases 3, 8 dan 9. Tambahan pula, antrakuinon ini mendorong
apoptosis melalui laluan bebas p53. Damnacanthal juga menurunkan Bcl-2 dan
meningkatkan aktiviti Bax dalam sel CEM-SS. Sifat antikanser damnacanthal ini
menjadikannya agen berpotensi untuk merawat leukemia T-limfoblastik.
Kata
kunci: Antikanser; apoptosis; CEM-SS; damnacanthal
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*Corresponding
author; email: latifahsy@upm.edu.my
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