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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 apoptosis in Caco-2 cells. a Flow cytometry analysis of APOPercentage™ dye staining after exposure of ethanol extract of P. pycnantha (P) treated with various concentrations for 24 h. b Cells were treated with increasing concentrations of compound C2 and C3, apoptosis was assessed by APOPercentage™ assay for 24 h as determined by flow cytometry. The results represent the mean ± SEM of three independent experiments
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Fig3: Effect of Pleiocarpa pycnantha and isolated compounds on apoptosis in Caco-2 cells. a Flow cytometry analysis of APOPercentage™ dye staining after exposure of ethanol extract of P. pycnantha (P) treated with various concentrations for 24 h. b Cells were treated with increasing concentrations of compound C2 and C3, apoptosis was assessed by APOPercentage™ assay for 24 h as determined by flow cytometry. The results represent the mean ± SEM of three independent experiments

Mentions: The effect of P. pycnantha extract P and compounds C2 and C3 on Caco-2 cell growth was assessed using the APOPercentage ™ dye which detect apoptosis at the stage of phosphatidylserine externalization and its specific for the quantitation of apoptosis. Caco-2 cells were treated for 24 h with different concentrations of P, C2 and C3 and stained with APOPercentage ™ dye and analysed by flow cytometry. The extract P and the tested compounds induced significant levels of apoptosis between (25–38 % for P, 5–23 % for C2 and 6–47 %) in Caco-2 cells (Fig.3a and b). The extract and compounds induce apoptosis in a dose-dependent manner. The cytotoxicity caused by the extract P and compounds C2 and C3 may be due to in part to anti-proliferative and pro-apoptotic effects.Fig. 3


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 apoptosis in Caco-2 cells. a Flow cytometry analysis of APOPercentage™ dye staining after exposure of ethanol extract of P. pycnantha (P) treated with various concentrations for 24 h. b Cells were treated with increasing concentrations of compound C2 and C3, apoptosis was assessed by APOPercentage™ assay for 24 h as determined by flow cytometry. 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

Fig3: Effect of Pleiocarpa pycnantha and isolated compounds on apoptosis in Caco-2 cells. a Flow cytometry analysis of APOPercentage™ dye staining after exposure of ethanol extract of P. pycnantha (P) treated with various concentrations for 24 h. b Cells were treated with increasing concentrations of compound C2 and C3, apoptosis was assessed by APOPercentage™ assay for 24 h as determined by flow cytometry. The results represent the mean ± SEM of three independent experiments
Mentions: The effect of P. pycnantha extract P and compounds C2 and C3 on Caco-2 cell growth was assessed using the APOPercentage ™ dye which detect apoptosis at the stage of phosphatidylserine externalization and its specific for the quantitation of apoptosis. Caco-2 cells were treated for 24 h with different concentrations of P, C2 and C3 and stained with APOPercentage ™ dye and analysed by flow cytometry. The extract P and the tested compounds induced significant levels of apoptosis between (25–38 % for P, 5–23 % for C2 and 6–47 %) in Caco-2 cells (Fig.3a and b). The extract and compounds induce apoptosis in a dose-dependent manner. The cytotoxicity caused by the extract P and compounds C2 and C3 may be due to in part to anti-proliferative and pro-apoptotic effects.Fig. 3

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