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Pleiocarpa pycnantha leaves and its triterpenes induce apoptotic cell death in Caco-2 cells in vitro.

Omoyeni OA, Hussein A, Meyer M, Green I, Iwuoha E - BMC Complement Altern Med (2015)

Bottom Line: The exposure of an ethanolic extract from the leaves of P. pycnantha (0.1-1000 μg/ml) and the isolated compounds C2 and C3 (6,25-100 μg/ml) to human colorectal cancer cells reduced the cell viability with an IC50 > 100, 40.9, 36.3 μg/ml for P, C2 and C3 respectively, after 24 h of incubation.Caspase 3 was also activated which is a hallmark of apoptosis.A further study with other cell lines is also recommended.

View Article: PubMed Central - PubMed

Affiliation: Department of Chemistry, University of the Western Cape, Bellville, South Africa. nikxyglo@yahoo.com.

ABSTRACT

Background: Recently, we reported that the crude fractions and pure triterpenes; ursolic acid (C1), 27-E and 27-Z p-coumaric esters of ursolic acid (C2, C3), together with a new triterpene 2,3-seco-taraxer-14-en-2,3-lactone [pycanocarpine (C4)] and its hydrolysed derivative - (2,3-seco-taraxen-4-hydroxy-14-en-2-oic acid) [pycanocarpene (C5)] from Pleiocarpa pycnantha leaves inhibit cell proliferation. However, there has not been any specific report on the use of Pleiocarpa pycnantha leaves and its constituents to kill colorectal adenocarcinoma cancer CaCo-2 cells. We performed in vitro study to evaluate the cytotoxic properties of the ethanolic extract of P. pycnantha P, compounds C2 and C3. A preliminary study of the potential mechanisms were also undertaken.

Methods: Cell viability was measured by WST-1 assay. The Apoptosis level was evaluated by staining with APOPercentage(™) dye and the induction of caspases 3/7 and 9 using Caspase-Glo(®) assays.

Results: The exposure of an ethanolic extract from the leaves of P. pycnantha (0.1-1000 μg/ml) and the isolated compounds C2 and C3 (6,25-100 μg/ml) to human colorectal cancer cells reduced the cell viability with an IC50 > 100, 40.9, 36.3 μg/ml for P, C2 and C3 respectively, after 24 h of incubation. The APOPercentage(TM) assay also showed a considerable increase in the percentage of apoptotic cells after 24 h; (25-38% for P, 5-23% for C2 and 6-47 % for C3). Caspase 3 was also activated which is a hallmark of apoptosis.

Conclusion: These findings suggest that the P. pycnantha and the isolated compounds induce cell apoptosis in human colorectal adenocarcinoma cells. A further study with other cell lines is also recommended.

No MeSH data available.


Related in: MedlinePlus

Effect of Pleiocarpa pycnantha and isolated compounds on the viability of Caco-2 cells. a Cells were treated with various concentrations of ethanol extract of P. pycnantha P and the relative cell viability was assessed by WST-1 assay for 24 h. b Cells were treated with various concentrations of compound C2 and C3, the relative cell viability was assessed by WST-1 assay for 24 h. The results represent the mean ± SEM of three independent experiments
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Fig2: Effect of Pleiocarpa pycnantha and isolated compounds on the viability of Caco-2 cells. a Cells were treated with various concentrations of ethanol extract of P. pycnantha P and the relative cell viability was assessed by WST-1 assay for 24 h. b Cells were treated with various concentrations of compound C2 and C3, the relative cell viability was assessed by WST-1 assay for 24 h. The results represent the mean ± SEM of three independent experiments

Mentions: P. pycnantha extract (P), isolated compounds (C2 and C3) decreased cell viability of Caco-2 cells. We examined the effect of P. pycnantha ethanolic extract and compounds C2 and C3 in cell viability using WST-1 assay. Caco-2 cells were treated with various concentrations of the extract and compounds C2 and C3, their viability was determined by the uptake of the formazan dye and expressed as percent of untreated control cells. The extract and the compounds induced a dose-dependent increase in viable formazan accumulating cells after the treatment (Fig. 2a and b). The 50 % growth inhibition concentration IC50 obtained after 24 h of incubation were > 100, 40.9, 36.3 μg/ml for P, C2 and C3 respectively.Fig. 2


Pleiocarpa pycnantha leaves and its triterpenes induce apoptotic cell death in Caco-2 cells in vitro.

Omoyeni OA, Hussein A, Meyer M, Green I, Iwuoha E - BMC Complement Altern Med (2015)

Effect of Pleiocarpa pycnantha and isolated compounds on the viability of Caco-2 cells. a Cells were treated with various concentrations of ethanol extract of P. pycnantha P and the relative cell viability was assessed by WST-1 assay for 24 h. b Cells were treated with various concentrations of compound C2 and C3, the relative cell viability was assessed by WST-1 assay for 24 h. The results represent the mean ± SEM of three independent experiments
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4499947&req=5

Fig2: Effect of Pleiocarpa pycnantha and isolated compounds on the viability of Caco-2 cells. a Cells were treated with various concentrations of ethanol extract of P. pycnantha P and the relative cell viability was assessed by WST-1 assay for 24 h. b Cells were treated with various concentrations of compound C2 and C3, the relative cell viability was assessed by WST-1 assay for 24 h. The results represent the mean ± SEM of three independent experiments
Mentions: P. pycnantha extract (P), isolated compounds (C2 and C3) decreased cell viability of Caco-2 cells. We examined the effect of P. pycnantha ethanolic extract and compounds C2 and C3 in cell viability using WST-1 assay. Caco-2 cells were treated with various concentrations of the extract and compounds C2 and C3, their viability was determined by the uptake of the formazan dye and expressed as percent of untreated control cells. The extract and the compounds induced a dose-dependent increase in viable formazan accumulating cells after the treatment (Fig. 2a and b). The 50 % growth inhibition concentration IC50 obtained after 24 h of incubation were > 100, 40.9, 36.3 μg/ml for P, C2 and C3 respectively.Fig. 2

Bottom Line: The exposure of an ethanolic extract from the leaves of P. pycnantha (0.1-1000 μg/ml) and the isolated compounds C2 and C3 (6,25-100 μg/ml) to human colorectal cancer cells reduced the cell viability with an IC50 > 100, 40.9, 36.3 μg/ml for P, C2 and C3 respectively, after 24 h of incubation.Caspase 3 was also activated which is a hallmark of apoptosis.A further study with other cell lines is also recommended.

View Article: PubMed Central - PubMed

Affiliation: Department of Chemistry, University of the Western Cape, Bellville, South Africa. nikxyglo@yahoo.com.

ABSTRACT

Background: Recently, we reported that the crude fractions and pure triterpenes; ursolic acid (C1), 27-E and 27-Z p-coumaric esters of ursolic acid (C2, C3), together with a new triterpene 2,3-seco-taraxer-14-en-2,3-lactone [pycanocarpine (C4)] and its hydrolysed derivative - (2,3-seco-taraxen-4-hydroxy-14-en-2-oic acid) [pycanocarpene (C5)] from Pleiocarpa pycnantha leaves inhibit cell proliferation. However, there has not been any specific report on the use of Pleiocarpa pycnantha leaves and its constituents to kill colorectal adenocarcinoma cancer CaCo-2 cells. We performed in vitro study to evaluate the cytotoxic properties of the ethanolic extract of P. pycnantha P, compounds C2 and C3. A preliminary study of the potential mechanisms were also undertaken.

Methods: Cell viability was measured by WST-1 assay. The Apoptosis level was evaluated by staining with APOPercentage(™) dye and the induction of caspases 3/7 and 9 using Caspase-Glo(®) assays.

Results: The exposure of an ethanolic extract from the leaves of P. pycnantha (0.1-1000 μg/ml) and the isolated compounds C2 and C3 (6,25-100 μg/ml) to human colorectal cancer cells reduced the cell viability with an IC50 > 100, 40.9, 36.3 μg/ml for P, C2 and C3 respectively, after 24 h of incubation. The APOPercentage(TM) assay also showed a considerable increase in the percentage of apoptotic cells after 24 h; (25-38% for P, 5-23% for C2 and 6-47 % for C3). Caspase 3 was also activated which is a hallmark of apoptosis.

Conclusion: These findings suggest that the P. pycnantha and the isolated compounds induce cell apoptosis in human colorectal adenocarcinoma cells. A further study with other cell lines is also recommended.

No MeSH data available.


Related in: MedlinePlus