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Advisor(s)
Abstract(s)
Beneficial effects have been attributed to Ganoderma lucidum including antioxidant, antitumor
and antibacterial properties [1-3]. Previous work from our group has identified a
methanolic extract (prepared at 25 °C) of this mushroom as being a potent modulator of
autophagy in a gastric cancer cell line (AGS) [4]. In the present work, the antitumor potential
and autophagy modulatory activity of a methanolic extract of G. lucidum, obtained
under cold extraction (-20°C), was further investigated. The chemical characterization
of the extract was previously published by some of the present authors [1]. Tumour cell
growth screening was carried out in four human tumor cell lines: AGS (gastric adenocarcinoma),
MCF-7 (breast adenocarcinoma), NCI-H460 (non-small cell lung cancer) and
HCT-15 (colorectal adenocarcinoma). The effect of the extract was further studied in
the most sensitive cells (AGS), particularly regarding effect in autophagy. The presence
of autophagossomes was observed following transfection of cells with a mCherry-LC3
expression vector and the levels of some autophagic proteins was analysed by Western
Blot. To confirm if the increase in LC3-II levels was a result from autophagy induction or
from a decrease in autophagic flux, cells were treated with the extract and with lysossomal
protease inhibitors (E-64d/pepstatin, to prevent formation of the autophagolysosome),
and the expression levels of autophagic proteins was analysed. We verified that
this extract provided a Gl50 of 66.59 iJQ/mL in the AGS cell line. In addition, treatment
with this extract (G150 concentration) caused an increase in the expression of LC3-II.
Additionally, the extract increased the formation of autophagossomes and when cells
were treated with the extract together with E-64d/pepstatin a further increase in the
LC3-IIIevels was verified, indicating that the extract caused an induction of autophagy.
Description
Keywords
Ganoderma lucidum Methanolic extract AGS Growth inhibition Autophagy
Citation
Reis, Filipa S.; Lima, Raquel T.; Morales, Patricia; Ferreira, Isabel C.F.R.; Vasconcelos, M. Helena (2014). A cold methanolic extract of Ganoderma lucidum induces autophagy in a gastric cancer cell line. In 62th International Congress and Annual Meeting of the Society for Medicinal Plant and Natural Product Research - GA2014. Guimarães