Sains Malaysiana 51(8)(2022): 2609-2617

http://doi.org/10.17576/jsm-2022-5108-20

Camellia sinensisand Phyllanthus amarusEthanol Extracts Induced Apoptosis and Cell Cycle Arrest on Human Leukemic Cell Lines

(Ekstrak Etanol Camellia sinensis dan Phylanthus amarus Mengaruh Apoptosis dan Hentian Kitaran Sel pada Titisan Sel Leukemia Manusia)

SYARATUL DALINA YUSOFF*, ENDANG KUMOLOSASI, MURSIDAH MD ALI, YAP REN QIAN, NORSYAHIDA MOHD. FAUZI & AFFIDAH SABRAN

Center for Drug & Herbal Development, Faculty of Pharmacy, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, 50300 Kuala Lumpur, Federal Territory, Malaysia

Received: 17 November 2021/Accepted: 28 January 2022

Abstract

Leukaemia is a heterogeneous hematologic malignancy characterized by unregulated proliferation of the early blood-forming cells which starts in bone marrow. The basic strategy of leukaemia therapy involves the induction of leukemic cells apoptosis. Research on natural products have shown that some plant derivatives have anticancer properties by inducing apoptosis of leukemic cells. Plants such as Camellia sinensis and Phyllanthus amarus are those that had gained a wide interest due to their anti-cancer effect. The aim of this study was to investigate the anti-cancer effects of C. sinensis and P. amarus extracts on human leukemic cell lines by analysing the cell cycle and determining the apoptotic state. The cell lines were treated with ethanolic plant extracts at the concentrations of 31.25 - 500 µg/mL for 24 h followed by MTT assay to determine the IC₅₀. The IC₅₀ of C. sinensis and P. amarus on the U937 cells were 170±10.39 and 210±6.78 µg/mL, respectively. Flow cytometric analysis of apoptosis using Annexin V/propidium iodide (PI) staining was also performed. C. sinensis extract at 170 µg/mL significantly increase apoptosis in U-937 (p<0.001), Jurkat (p<0.05) and K-562 cells (p<0.01) when compared to untreated cells. Meanwhile, P. amarus extract at 210 µg/mL significantly induced apoptosis in both U937 and K562 cells (p<0.05) but not Jurkat cells and caused cell cycle arrest at S phase in U-937 cells (p<0.001) and at G0/G1 in K652 cells (p<0.05) when compared to control. Based on the findings, both C. sinensis and P. amarus extracts showed potential in inducing apoptosis in human leukemic cell lines. In addition, P. amarus has the capability to disrupt cell cycle.

Keywords: Apoptosis; Camellia sinensis; cell cycle arrest; leukemia; Phyllanthus amarus

Abstrak

Leukemia adalah malignansi hematologi heterogen, dicirikan oleh pemproliferatan sel pembentuk darah awal secara tidak terkawal, yang bermula di dalam sumsum tulang. Sel leukemia mengalami apoptosis teraruh yang merupakan strategi asas dalam terapi leukemia. Kajian yang melibatkan produk semula jadi ada menunjukkan ciri-= antikanser terhadap sel leukemia melalui apoptosis teraruh. Tumbuhan seperti Camellia sinensis dan Phyllanthus amarus adalah antara yang mendapat perhatian meluas kerana mempunyai ciri antikanser. Matlamat penyelidikan ini adalah untuk mengkaji kesan antikanser ekstrak C. sinensis dan P. amarus terhadap pelbagai jenis sel leukemia manusia dengan menganalisis kitaran sel dan menentukan keadaan apoptosis. Titisan sel dirawat dengan ekstrak etanol tumbuhan pada kepekatan 31.25 - 500 µg/mL selama 24 jam dan diikuti dengan ujian MTT untuk menentukan IC₅₀. IC₅₀ C. sinensis dan P. amaruspada sel U937 masing-masing adalah 170±10.39 dan 210±6.78 µg/mL. Analisis apoptosis sitometri aliran menggunakan pewarnaan Annexin V/propidium iodida (PI) juga dilakukan. Ekstrak C. sinensispada 170 µg/mL meningkatkan apoptosis pada sel U-937 (p<0,001), Jurkat (p<0,05) dan K-562 (p<0,01) secara signifikan jika dibandingkan dengan sel yang tidak dirawat. Sementara itu, ekstrak P. amarus pada 210 µg/mL telah menyebabkan apoptosis pada sel U937 dan K562 (p<0.05) secara signifikan tetapi tidak pada sel Jurkat serta menyebabkan hentian kitaran sel pada fasa S pada sel U-937 dengan peratusan 55.71±2.06% (p<0.001) dan pada fasa G0/G1 pada sel K652 dengan peratusan 56.77±4.69% (p<0.05) jika dibandingkan dengan kawalan. Berdasarkan penemuan ini, kedua-dua ekstrak C. sinensis dan P. amarus menunjukkan potensi dalam mengaruh apoptosis terhadap sel leukemia manusia dan P. amarus berupaya untuk mengganggu kitaran sel.

Kata kunci: Apoptosis; Camellia sinensis; hentian kitaran sel; leukemia; Phyllanthus amarus

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^*Corresponding author; email: dalina.yusoff@ukm.edu.my