Antiproliferative activity and induction of apoptosis by Annona muricata (Annonaceae) extract on human cancer cells
1
Department of Physiological Sciences and Biochemistry, Faculty of
Medicine and Biomedical Sciences, University of Yaoundé I, PO Box 1364,
Yaoundé, Cameroon
2 Cancer Pharmacology Division, Indian Institute of Integrative Medicine, 180001, Canal Road, 18001 Jammu, India
3 Department of Biochemistry and Molecular Biology, University of Buea, Buea, Cameroon
4 Department of Biochemistry, Faculty of Sciences, University of Yaoundé I, PO Box 812, Yaoundé, Cameroon
2 Cancer Pharmacology Division, Indian Institute of Integrative Medicine, 180001, Canal Road, 18001 Jammu, India
3 Department of Biochemistry and Molecular Biology, University of Buea, Buea, Cameroon
4 Department of Biochemistry, Faculty of Sciences, University of Yaoundé I, PO Box 812, Yaoundé, Cameroon
BMC Complementary and Alternative Medicine 2014, 14:516
doi:10.1186/1472-6882-14-516
The electronic version of this article is the complete one and can be found online at: http://www.biomedcentral.com/1472-6882/14/516
The electronic version of this article is the complete one and can be found online at: http://www.biomedcentral.com/1472-6882/14/516
| Received: | 3 June 2014 |
| Accepted: | 9 December 2014 |
| Published: | 24 December 2014 |
© 2014 Pieme et al.; licensee BioMed Central.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
Abstract
Background
Annona muricata (A. muricata) is widely distributed in Asia, Africa and South America. Different parts of this
plant are used to treat several diseases in Cameroon. The aim of this study is to
determine the in vitro anti-proliferative effects and apoptotic events of A. muricata extracts on HL-60 cells as well as to quantify its phenols content.
Methods
The cell viability was measured by using 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium
bromide (MTT) assay while the changes in morphology of HL-60 cells, membrane mitochondrial
potential (MMP) and the cell cycle were used for assessment apoptosis induction.
Results
The results show that the concentration of phenols, flavonoids and flavonols in the
extracts varied depending on the part of the plant. All the extracts tested inhibited
the proliferation of HL-60 cells in a concentration dependent manner with IC50 varied from 6–49 μg/mL. The growth inhibition of the cells by extracts was associated
with the disruption of MMP, reactive oxygen species (ROS) generation and the G0/G1
cell arrest.
Conclusion
These findings suggest that the extracts from A. muricata have strong antiproliferation potential and can induce apoptosis through loss of
MMP and G0/G1 phase cell arrest.
Keywords:
Apoptosis; Membrane mitochondrial potential; Antiproliferative; Cell cycle; A. muricataBackground
Chemopreventive properties have long been attributed to polyphenolic compounds present
in the human diet. The interest on these natural substances is increasing because
of their higher potential sources of anticancer compounds. Plants and plant-based
medicaments are used as the basis of many modern pharmaceuticals industries today
for the treatment of our various ailments [1]. According to world health organization (WHO), more than 80% of the total world’s
population depends on the traditional medicines to satisfy their primary health care
needs. Several phytochemical molecules from natural products capable of exerting a
physiologic action on the human body were studied and characterized. These bioactive
compounds such as alkaloids, flavonoids, tannins and phenols were considered to be
most important. The phytochemical research that has been done based on the ethno pharmacological
information forms the effective approach in the discovery of new anti-infective agents
from higher plants [2].
Annona muricata L (A. muricata) commonly known as graviola, soursop or corossol, belongs to the Annonaceae family.
It is a widespread small tree and has its native in Central America [3]. It is a typical tropical tree with heart shaped edible fruits and widely distributed
in most of tropical countries. All parts of A. muricata tree are used in natural medicine in the tropic including the twigs, leaf, root,
fruit and seeds. Generally, the fruit and fruit juice are taken to eliminate worms
and parasites, cool fever, increase mother’s milk after child birth, and as an astringent
for diarrhea and dysentery [4]. The crushed seeds are used against internal and external parasites, head lice and
warms. The twigs, leaf are considered sedative and antispasmodic [4]. A decoction of A. muricata leaf is used to kill bed bug and head lice to reduce fever. For the latter it can
have the same effect taken orally or added to bathing water [5]. The creamy and delectable flesh of the fruit consist of 80% water, 1% protein, 18%
carbohydrates and fair amount of vitamins B, B2 and C, potassium and dietary fiber
[6]. The leaf are lanceolate with glossy and dark green in color had been traditionally
used to treat headaches, hypertension, cough, asthma and used as antispasmodic, sedative
and nervine for heart condition [7,8]. Previous reports have demonstrated that the leaf, twigs, root, stem, and fruit seed
extracts of A. muricata have several biological activities such as anti-bacterial [9], antifungal [10] and anti-malarial [11]. Its leaf extract were also found to possess antioxidant [12] and molluscicidal properties [13]. Recently, it has also been reported to exhibit anti-inflammatory and analgesic effects
[14], cytotoxicity and apoptosis inducing activities on T47D breast cancer [15], antiviral activity [16] and antidiabetic activity. Phytochemical investigation of the leaf of A. muricata showed the presence of alkaloids [17], essential oils [18] and acetogenins [19]. These acetogenins demonstrated to be selectively toxic against various types of
the cancerous cells without harming healthy cells [20]. Acetogenin 1 was reported to exhibit cytotoxic activities against the human pancreatic
tumor cell line (PACA-2), human prostate adenocarcinoma (PC-3) and human lung carcinoma
(A-549), while Acetogenin 2 was reported to exhibit cytotoxicity against human hepatoma
carcinoma cell line (Hep G2) [21]. Seven isoquinoline alkaloids including reticuline, coclaurine, coreximine, atherosperminine,
stepharine, anomurine and anomuricine have been isolated from the leaves, root and
stem barks of A. muricata[22]. The essential oil of the fresh fruit pulp of A. muricata contains 2-hexenoic acid methyl ester (23.9%), 2-hexenoic acid ethyl ester (8.6%),
2-octenoic acid methyl ester (5.4%), 2-butenoic acid methyl ester (2.4%), β-caryophyllene
(12.7%), 1,8-cineole (9.9%), linalool (7.8%), α-terpineol (2.8%), lialyl propionate
(2.2%) and calarence (2.2%) [23]. Therefore, we attempted to investigate the growth-inhibitory and apoptotic effects
of extracts from leaf, twigs and roots from A. muricata against Human promyelocytic leukemia (HL-60 cells).
