Citation
Lam, Tsuey Peng
(2007)
Morphological and Molecular Characterisation of Ethanolic Neem (Azadirachta Indica) Leaf Extract In an in Vivo Breast Cancer Model.
Masters thesis, Universiti Putra Malaysia.
Abstract
Breast cancer is the commonest cause of cancer death in women worldwide
and Malaysia in all ethnic groups and all age groups. Neem's (Azadirachta
indica) ability as a medicinal herb is traced as far back as 4500 years ago.
Some of the impressive therapeutic qualities have been discovered such as
anti-viral, anti-microbial, anti-inflammatory, anti-tumour, anti-bacterial,
anti-fungal and anti-hyperglycemic; however the anticancer effect of ethanolic
Neem leaves extract against breast cancer has not been documented.
Besides this, Neem was found to induce apotosis in MCF-7 breast cancer cell
line in local study recently. Thus, this study was done to evaluate the effect of
ethanolic Neem leaves extract as apoptosis inducer in in vivo 4Tl breast
cancer model. Two different concentrations of Neem, 250 mgkg and 500 mglkg were tested on 4T1 breast cancer model. The 4T1 breast cancer
models were evaluated by light microscopy, transmission electron microscopy
for morphological changes, TUNEL assay for apoptotic cell labeling and in
situ RT-PCR for c-myc, c-erbB2 and c-fos oncogene expressions. All
treatment groups exhibited a higher incidence of apoptosis compared to
untreated group from morphological analysis and TUNEL assay. The
cancerous mice treated with both different concentration of Neem showed
significantly higher value (p<0.05) in mean body weight, mean apoptotic index
and mean apoptotic score compared to the control group. At the same time
both group were showing a significantly lower value of mean mitotic index in
histological evaluation. The mean tumour volume and mass proved that
there was evidence of tumour regression in Neem treated mice. However, the
overall observation showed that 500 mglkg of Neem has more significant
effect (p <0.05) of inducing apoptosis in the 4T1 breast cancer cells compared
to 250 mgkg of Neem. Furthermore, the 500 mgkg Neem concentration has
significantly lengthened the mean survival time by 44.62% in the 4T1 breast
cancer model (p <0.05). Neem 500 mglkg group also showed a better
suppression of c-myc, c-erbB2 and c-fos oncogenes expression in mean
distribution and intensity score (pc 0.05) in the 4T1 breast cancer model. By
considering all the three down regulated oncogenes (c-myc, c-erbB2 and
c-fos) under effect of Neem 500 mgkg together, it becomes clearer that
Neem 500 mglkg was effective in inducing apoptosis in the 4T1 breast cancer model. In conclusion, the Neem 500 mglkg treatment was effective in inducing
cell death via apoptosis and regulates cell proliferation in 4T1 breast cancer
model. Its effectiveness was proportional to the concentration of Neem
treatment given.
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