Annona muricata leaves induce G cell cycle arrest and apoptosis

through mitochondria-mediated pathway in human HCT-116 and

HT-29 colon cancer cells
I.

II.

III.

IV.

V.

VI.

Product / Ingredient Studied

a.
Annona muricata L. (Annona muricata) as a member of the
Annonaceae family, the custard apple family, is a small tropical tree
widely cultivated throughout the tropical countries. Annona species
such as Annona squamosa and Annona reticulata are well known to
have traditional application in the tropics
b. The popular fruit tree of Annona muricata known also as graviola or
soursop has extensive ethnobotonical uses, such as sedative,
astringent,piscicide,insecticide,vermifuge,hypotensive,antiparasiticand
antispasmodic.
c. Furthermore, it is traditionally used for the treatment of fevers,
asthma, pain, coughs, wound and skin remedies
d. Annona muricata was described as the cancer killer due to its
remarkable cytotoxicity against various cancer cell lines. The seeds
and leaves of Annona muricata have been widely used by the natives
in South America for the treatment of cancer and tumors.
Indication
a. To establish the possible apoptotic pathways induced in HT-29 and
HCT-116 colon cancer cells by ethyl acetate extract of Annona
muricata leaves.
Type of Claim
a. The stems, barks and leaves of Annona muricata revealed noteworthy
antiproliferative effects against cancer cells without affecting normal
cells
Dosage and Administration
a. The antiproliferative potential of the three isolated extracts was
expressed as IC50 value, the concentration of plant extract that caused
50% inhibition of cell growth which was calculated based on the
percentage of cell viability. As ethyl acetate extract of Annona muricata
leaves (EEAM) showed the strongest IC50, which was used to continue
the study.
Type of Study (Human or Animal)
a. Human cells
b. CD841 (normal human colon epithelial cells)
c. HT-29 and HCT116 (human colon cancer cells)
Study Design (Experimental or Observational)
a. Experimental in vitro
b. CCD841 (normal human colon epithelial cells), HT-29 and HCT116
(human colon cancer cells) were obtained from the American Type Cell
Collection (ATCC, Manassas, VA, USA). The cells were maintained in
RPMI-1640 medium (Sigma, St. Louis, MO, USA) supplemented with

VII.
VIII.
IX.

X.
XI.
XII.

XIII.

10% FBS (PAA Lab, Pasching, Austria), 50 g/ml amphotericin B (PAA

Lab) and 100 U/ml penicillin (PAA Lab) in a humidied atmosphere with
5% CO in the air at 37 1C. Cells in the exponential growth phase were
collected for the following experiments. The negative control for all the
assays was represented by the untreated medium containing vehicle
DMSO (0.1%).
Study Population
a. Colon cancer
Duration of the Study
a. Incubated for 72 h.
Study End Points
a. Cell viability was evaluated by using the 3-(4,5-dimethylthiazol-2-yl)2,5-diphenyltetrazolium (MTT) assay.
b. In brief, cells (5 x 10 4 cells/ml) were treated with three isolated
extracts at different concentrations in 96-well plate and incubated for
72 h.
Ethics Committee Approval
a. Not mentioned within the article.
Limitation of the Study
a. Not stated within the article.
Study Results
a. EEAM exerted signicant cytotoxic effects on HCT-116 and HT-29 cells
as determined by MTT and LDH assays.
b. After 24 h treatment, EEAM exhibited the IC value of 11.4371.87 mg/ml
and 8.9871.24 mg/ml against HT-29 and HCT-116 cells, respectively.
Flow cytometric analysis demonstrated the cell cycle arrest at G1 50
phase and phosphatidylserine externalization conrming the induction
of apoptosis.
c. EEAM treatment caused excessive accumulation of ROS followed by
disruption of MMP, cytochrome c leakage and activation of the initiator
and
executioner
caspases
in
both
colon
cancer
cells.
Immunouorescence analysis depicted the up-regulation of Bax and
down-regulation of Bcl-2 proteins while treated with EEAM.
d. Furthermore, EEAM conspicuously blocked the migration and invasion
of HT-29 and HCT-116 cells.
Source of Evidence:
a. Moghadamtousi, S.Z., et al., Annona muricata leaves induce G1 cell
cycle arrest and apoptosis through mitochondria-mediated pathway in
human....
Journal
of
Ethnopharmacology
(2014),
http://dx.doi.org/10.1016/j.jep.2014.08.011i