Muhamad, Nor ‘Atiqah
(2018)
Clearance of apoptotic cells by macrophage in MCF-7 cell line treated with methanol extract of centella asiatica (MECA).
Masters thesis, Universiti Sains Malaysia.
Abstract
Breast cancer is one of the major contributions in non-communicable diseases
worldwide. To date, much evidence has shown that resistance to apoptosis by cancer
cells and inefficient efferocytosis by phagocytic cells in cancer patients foster the
development of other diseases in cancer patients, such as autoimmune diseases.
Previous studies have shown that Centella asiatica (C. asiatica) extract induced
apoptosis in cancer cell line but there was a lack of study regarding efferocytosis against
cancer cells. C. asiatica has the potential to be an alternative treatment to induce
apoptosis and efferocytosis by phagocytic cells in cancer patients. The anti-proliferative
activity of methanol extract of C. asiatica (MECA) against breast cancer cell line
(MCF-7) was obtained by performing methylene blue assay (MBA). In this method,
DMSO served as a negative control while tamoxifen served as a positive control. After
that, the mode of cell death in MECA-treated cells was determined by Hoechst 33342
staining and was further investigated by flow cytometry analysis. The findings on the
MBA suggested that MECA was cytotoxic on MCF-7 and not toxic on normal cells
(NIH3T3). Furthermore, MCF-7 treated with MECA undergone increased apoptosis
indicated by brighter fluorescence, alteration of nuclear morphology and DNA
fragmentation observed in Hoechst staining. To study efferocytosis activity by
macrophages (J774A.1), MCF-7 cells were cultivated in J774A.1 cells with multiplicity
of infection (MOI) of 1:2. After treatment with MECA, flow cytometry data showed a
significant increase of late apoptotic cells due to apoptosis of J774A.1 after engulfing
MCF-7 cells. In conclusion, MECA was able to improve the activities of apoptosis and
efferocytosis in MCF-7 cells, thus suggesting a promising alternative treatment for
breast cancer.
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