Loading...
Research Project
SMALL MOLECULES AS APOPTOSIS INDUCERS IN CANCER CELLS: INVESTIGATION OF THE MECHANISM OF ACTION
Funder
Authors
Publications
Synthesis of aminodiarylamines in the thieno[3,2-b]pyridine series and effects on tumor cell growth inhibition, cell cycle and apoptosis
Publication . Calhelha, Ricardo C.; Ferreira, Isabel C.F.R.; Abreu, Rui M.V.; Vale-Silva, Luís A.; Pinto, Eugénia; Lima, Raquel T.; Alvelos, M. Inês; Vasconcelos, M. Helena; Queiroz, Maria João R.P.
Several series of compounds that include the
thienopyridine scaffold have been reported as
inhibitors of known cancer therapeutic targets or
as inhibitors of cell proliferation in tumor cell lines
[1,2]. Our research group has already
synthesized several thieno[3,2-b]pyridine
derivatives by Pd-catalyzed C-C (Suzuki and
Sonogashira) and C-N (Buchwald-Hartwig)
couplings and some of them have presented
tumor cell growth inhibitory activity in cell lines [3-
5].
A cold methanolic extract of Ganoderma lucidum induces autophagy in a gastric cancer cell line
Publication . Reis, Filipa S.; Lima, Raquel T.; Morales, Patricia; Ferreira, Isabel C.F.R.; Vasconcelos, M. Helena
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.
An aqueous extract of tuberaria lignosa inhibits cell growth, alters the cell cycle profile, and induces apoptosis of NCI-H460 tumor cells
Publication . Pereira, Joana M.; Lopes-Rodrigues, Vanessa; Xavier, Cristina; Lima, M. João; Lima, Raquel T.; Ferreira, Isabel C.F.R.; Vasconcelos, M. Helena
Tuberaria lignosa (Sweet) Samp. is found in European regions, and has antioxidant properties due to its composition in ascorbic acid and phenolic compounds. Given its traditional use and antioxidant properties, the tumor cell growth inhibitory potential of aqueous extracts from T. lignosa (prepared by infusion and decoction) was investigated in three human tumor cell lines: MCF-7 (breast adenocarcinoma), NCI-H460 (non-small cell lung cancer), and HCT-15 (human colorectal adenocarcinoma). Both extracts inhibited the growth of these cell lines; the most potent one being the T. lignosa extract obtained by infusion in the NCI-H460 cells (GI50 of approximately 50 μg/mL). Further assays were carried out with this extract in NCI-H460 cells. At 100 μg/mL or 150 μg/mL it caused an increase in the percentage of cells in the G0/G1 phase and a decrease of cells in S phase of the cell cycle. Additionally, these concentrations caused an increase in the percentage of apoptotic cells. In agreement, a decrease in total poly (ADP-ribose) polymerase (PARP) and pro-caspase 3 levels was found. In conclusion, the T. lignosa extract obtained by infusion was more potent in NCI-H460 cells, altering the cell cycle progression and inducing apoptosis. This work highlights the importance of T. lignosa as a source of bioactive compounds with tumor cell growth inhibitory potential.
Cordyceps militaris (L.) Link fruiting body reduces the growth of a non-small cell lung cancer cell line by increasing cellular levels of p53 and p21
Publication . Bizarro, Ana; Ferreira, Isabel C.F.R.; Soković, Marina; Van Griensven, Leo J.L.D.; Sousa, Diana; Vasconcelos, M. Helena; Lima, Raquel T.
Cordyceps militaris (L.) Link, an edible entomopathogenic fungus widely used in traditional Chinese medicine, has numerous potential medicinal properties including antitumor activity. The methanolic extract of C. militaris fruiting body was recently shown to have tumor cell growth inhibitory activity in several human tumor cell lines. Nonetheless, the mechanism of action involved is still not known. This work aimed at further studying the effect of the methanolic extract of C. militaris regarding its antitumor mechanism of action, using the non-small cell lung cancer cell line (NCI-H460) as a model. Results showed that treatment with the extract decreased cellular proliferation, induced cell cycle arrest at G0/G1 and increased apoptosis. In addition, the extract increased the levels of p53 and p21. Moreover, an increase in p-H2A.X and 53BP1 levels, together with an increase in the number of 53BP1 foci/cell (all indicative of DNA damage), were also observed after treatment with the extract. This work suggests that this extract affected NCI-H460 cellular viability through a mechanism involving DNA damage and p53 activation. This further supports the potential of this extract as a source of bioactive compounds, which may be used in anticancer strategies.
Cordyceps militaris (L.) link fruiting body reduces NCI-H460 cellular viability through a mechanism involving p53 and p21
Publication . Bizarro, Ana; Ferreira, Isabel C.F.R.; Soković, Marina; Van Griensven, Leo J.L.D.; Sousa, Diana; Vasconcelos, M. Helena; Lima, Raquel T.
Mushroom extracts are recognized by their numerous potential medicinal properties.
Recently, a methanolic extract from Cordyceps militaris (L.) Link (an edible
entomopathogenic fungus widely used in traditional Chinese medicine) has been
shown to inhibit cell growth of several human tumour cell lines [1,2]. However, its
mechanism of action remained unknown.
The aim of the present work was to study the antitumour mechanism of action of the
methanolic extract of C. militaris, in the NCI-H460 cell line which is representative of
non-small cell lung cancer.
Results showed that the extract reduced viable cell number (observed with the trypan
blue exclusion assay) by: i) decreasing cellular proliferation (analysed with the BrdU
incorporation assay), ii) inducing cell cycle arrest at GO/GI phase (analysed by flow
cytometry following propidium iodide-PI labelling) and iii) increasing apoptosis
(analysed by flow cytometry following Annexin V-FITC and PI labelling). In addition,
results also showed that treatment with the extract increased the cellular levels of p53
and p21. Moreover, this study also showed evidences of DNA damage caused by this
extract, since an increase in the levels ofp-H2A.X and 53BP1 were observed, together
with an increase in the number of 53BP1 foci/cell.
In conclusion, this extract reduced NCI-H460 cellular viability, possibly through a
mechanism which involves DNA damage and p53.
Organizational Units
Description
Keywords
Contributors
Funders
Funding agency
Fundação para a Ciência e a Tecnologia
Funding programme
Funding Award Number
SFRH/BPD/68787/2010