Volume 168, 20 June 2015, Pages 50–60
Glycosmis pentaphylla (Retz.) DC arrests cell cycle and induces apoptosis via caspase-3/7 activation in breast cancer cells
Abstract
Ethnopharmacological relevance
Glycosmis pentaphylla
(Retz.) DC belonging to the family Rutaceae has been traditionally used
for the treatment of rheumatism, anaemia, jaundice, skin diseases,
bronchitis etc. The plant is traditionally considered as anti-cancer
medicine and used by the healers of Bangladesh to treat all types of
cancers. Perhaps the key to many of its medicinal applications is its
inherent anti-inflammatory property.
Aim of the study
The present study is aimed at evaluating the effect of various fractions of G. pentaphylla (Retz.) DC leaves on the cell cycle and apoptosis of breast cancer cells viz. MCF-7 and MDA-MB-231.
Materials and methods
Various extracts and fractions of the leaves of G. pentaphylla
(Retz.) DC were studied for their cytotoxicity with the help of
Sulforhodamine B assay, in MCF-7, MDA-MB-231 and Vero cell lines. The
most active fractions were studied for their effect on the cell cycle of
MCF-7 and MDA-MB-231 cells. Apoptotic studies were done using Hoechst
staining, DNA fragmentation, Annexin V staining and caspase-3/7
activation assay in breast cancer cells. HPLC and HPTLC profiling of the
active fractions were done.
Results
HPTLC
and HPLC profiling revealed the presence of lupeol, chrysin, quercetin,
β-sitosterol and kaempferol as components in active fractions. Lupeol
and chrysin are being reported in this plant for the first time. The
studies showed that the selected fractions possess cell cycle inhibitory
and apoptosis inducing effect on both MCF-7 and MDA-MB-231 cells.
Apoptotic effect of the fractions on MCF-7 and MDA-MB-231 cells may be
through the mitochondrial pathway by the activation of caspase-3/7.
Graphical abstract
Abbreviations
- SRB Assay, sulforhodamine B assay;
- DMEM, Dulbecco’s minimum essential media;
- FBS, fetal bovine serum;
- PI, propidium iodide;
- NP-40, nonidet P-40;
- PBS, phosphate buffer saline;
- DPPH assay, 1,1-diphenyl 1-2-picryl-hydrazyl assay;
- ABTS assay, 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) assay;
- 2,4-DNPH, 2,4-dinitrophenylhydrazine;
- OD, optical density;
- PBS, phosphate buffer saline;
- 7-AAD, 7-aminoactinomycin D;
- AUC, area under the curve;
- S.E.M., standard error of mean;
- RT, retention time.
Keywords
- Glycosmis pentaphylla (Retz.) DC;
- Apoptosis;
- Cell cycle arrest;
- Caspase-3/7 activation;
- Annexin V staining;
- HPLC
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