Phytochemical screening and natural killer cells immunomodulation effects of pereskia bleo leaves extract on cervical cancer cells
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Date
2020-08
Authors
Salleh, Siti Farhanah Mohd
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Abstract
Pereskia bleo is a leafy and edible plant, locally known as 'Pokok Jarum Tujuh
Bilah" which has anti-cancer properties. This study purposed to elucidate the
underlying mechanism of this plant as anti-cancer in inducing cell death as well as to
evaluate its immunostimulatory effects on Natural Killer cells (NK cells) as a potential
additional anti-cancer effect. In this study, the leaves of P. bleo were extracted using
propenyl) and hexadecanoic acid. PBEA exhibited the lowest IC50 value (14.37 ± 8.40
μg/ml) indicated the strongest cytotoxic effect selectively on cervical cancer cells
(HeLa). The cell cycle analysis showed inhibition of cell proliferation at G0/G1 phase
in PBEA treated HeLa cells as evidenced by a significant accumulation of the cells at
this phase (P<0.05). Morphological examination on PBEA treated HeLa cell showed
the presence of fragmented nuclei and condensation of chromatin while apoptosis was
detected in the Annexin V/PI assay. Analysis of apoptotic proteins revealed a
significant upregulation of pro-apoptotic proteins (Bax, p53 and caspase-3) while
downregulation of anti-apoptotic protein Bcl-2 (P<0.05) in PBEA treated HeLa cells.
Meanwhile, NK cells proliferation at 24 h was found significantly increased compared
to 48 h and 72 h of PBEA treatment (P<0.05). Apoptosis of HeLa cells was markedly
increased in PBEA treated NK cells from cancer patients. This extract also enhanced
granzyme B and IFN- epression in NK cells from cancer paiens. Thus our findings
demonstrated that PBEA induced cell death in the cervical cancer cells (HeLa) and
stimulate activation of NK cells from cervical cancer patients which enhanced
cytotoxic effect against HeLa cells. These results provide some insight into the
effectiveness of P. bleo as a potential chemopreventive agent which open up for further
studies.
different techniques and solvent polarities, and subsequently subjected to GC-MS
analysis. The extracts were tested for its cytotoxic effects on HeLa, MDA-MB-231,
SW480 and NIH/3T3 cell lines using MTT assay. The most cytotoxic extract and its
corresponding cancer cell lines were investigated for their cell death induction through
cell cycle arrest, Annexin V/PI assay and measurement of apoptotic proteins using
flow cytometry. NK cells were exposed to different concentrations of ethyl acetate
extract of P. bleo leaves (PBEA) and its proliferation rate was determined via MTT
assay. NK cells from healthy individuals and cervical cancer patients were treated with
14.4 μg/ml of PBEA and co-cultured with target cells for 24 h to evaluate its cytotoxic
activity. Target cells death was identified by flow cytometry while ELISA assay was
performed to determine the production of perforin, granzyme B, IFN- and IL-2.
Results showed the presence of terpenoids, sterols, alkaloids, flavonoids, phenols, fatty
acids and vitamin E in the extracts of P. bleo leaves together with new compounds
namely (-)-Loliolide, neophytadiene, -ocopherol, -tocopherol, squalene, 4H-Pyran-
4-one,2,3-dihydro-3,5-dihydroxy-6-methyl, 4-vinyl-syringol, phenol,2-methoxy-4-(1-
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Keywords
anti-cancer