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Events associated with apoptotic effect of p -Coumaric acid in HCT-15 colon cancer cells

AIM: To investigate the events associated with the apoptotic effect of p -Coumaric acid, one of the phenolic components of honey, in human colorectal carcinoma (HCT-15) cells. METHODS: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltertazolium- bromide assay was performed to determine the antiproliferativ...

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Main Authors: Jaganathan, Saravana Kumar, Supriyanto, Eko, Mandal, Mahitosh
Format: Article
Language:English
Published: Baishideng Publishing Group Co. Limited 2013
Subjects:
R Medicine
Online Access:http://eprints.utm.my/id/eprint/50550/1/EkoSupriyanto2013_Eventsassociatedwithapoptotic.pdf
http://eprints.utm.my/id/eprint/50550/
http://dx.doi.org/10.3748/wjg.v19.i43.7726
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spelling my.utm.505502018-10-14T08:37:02Z http://eprints.utm.my/id/eprint/50550/ Events associated with apoptotic effect of p -Coumaric acid in HCT-15 colon cancer cells Jaganathan, Saravana Kumar Supriyanto, Eko Mandal, Mahitosh R Medicine AIM: To investigate the events associated with the apoptotic effect of p -Coumaric acid, one of the phenolic components of honey, in human colorectal carcinoma (HCT-15) cells. METHODS: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltertazolium- bromide assay was performed to determine the antiproliferative effect of p -Coumaric acid against colon cancer cells. Colony forming assay was conducted to quantify the colony inhibition in HCT 15 and HT 29 colon cancer cells after p -Coumaric acid treatment. Propidium Iodide staining of the HCT 15 cells using flow cytometry was done to study the changes in the cell cycle of treated cells. Identification of apoptosis was done using scanning electron microscope and photomicrograph evaluation of HCT 15 cells after exposing to p -Coumaric acid. Levels of reactive oxygen species (ROS) of HCT 15 cells exposed to p -Coumaric acid was evaluated using 2', 7'-dichlorfluorescein- diacetate. Mitochondrial membrane potential of HCT-15 was assessed using rhodamine-123 with the help of flow cytometry. Lipid layer breaks associated with p -Coumaric acid treatment was quantified using the dye merocyanine 540. Apoptosis was confirmed and quantified using flow cytometric analysis of HCT 15 cells subjected to p -Coumaric acid treatment after staining with YO-PRO-1. RESULTS: Antiproliferative test showed p -Coumaric acid has an inhibitory effect on HCT 15 and HT 29 cells with an IC50 (concentration for 50% inhibition) value of 1400 and 1600 μmol/L respectively. Colony forming assay revealed the time-dependent inhibition of HCT 15 and HT 29 cells subjected to p -Coumaric acid treatment. Propidium iodide staining of treated HCT 15 cells showed increasing accumulation of apoptotic cells (37.45 ± 1.98 vs 1.07 ± 1.01) at sub-G1 phase of the cell cycle after p -Coumaric acid treatment. HCT-15 cells observed with photomicrograph and scanning electron microscope showed the signs of apoptosis like blebbing and shrinkage after p -Coumaric acid exposure. Evaluation of the lipid layer showed increasing lipid layer breaks was associated with the growth inhibition of p -Coumaric acid. A fall in mitochondrial membrane potential and increasing ROS generation was observed in the p -Coumaric acid treated cells. Further apoptosis evaluated by YO-PRO-1 staining also showed the timedependent increase of apoptotic cells after treatment. CONCLUSION: These results depicted that p -Coumaric acid inhibited the growth of colon cancer cells by inducing apoptosis through ROS-mitochondrial pathway Baishideng Publishing Group Co. Limited 2013 Article PeerReviewed application/pdf en http://eprints.utm.my/id/eprint/50550/1/EkoSupriyanto2013_Eventsassociatedwithapoptotic.pdf Jaganathan, Saravana Kumar and Supriyanto, Eko and Mandal, Mahitosh (2013) Events associated with apoptotic effect of p -Coumaric acid in HCT-15 colon cancer cells. World Journal of Gastroenterology, 19 (43). pp. 7726-7734. ISSN 1007-9327 http://dx.doi.org/10.3748/wjg.v19.i43.7726 DOI: 10.3748/wjg.v19.i43.7726
institution Universiti Teknologi Malaysia
building UTM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Teknologi Malaysia
content_source UTM Institutional Repository
url_provider http://eprints.utm.my/
language English
topic R Medicine
spellingShingle R Medicine
Jaganathan, Saravana Kumar
Supriyanto, Eko
Mandal, Mahitosh
Events associated with apoptotic effect of p -Coumaric acid in HCT-15 colon cancer cells
description AIM: To investigate the events associated with the apoptotic effect of p -Coumaric acid, one of the phenolic components of honey, in human colorectal carcinoma (HCT-15) cells. METHODS: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltertazolium- bromide assay was performed to determine the antiproliferative effect of p -Coumaric acid against colon cancer cells. Colony forming assay was conducted to quantify the colony inhibition in HCT 15 and HT 29 colon cancer cells after p -Coumaric acid treatment. Propidium Iodide staining of the HCT 15 cells using flow cytometry was done to study the changes in the cell cycle of treated cells. Identification of apoptosis was done using scanning electron microscope and photomicrograph evaluation of HCT 15 cells after exposing to p -Coumaric acid. Levels of reactive oxygen species (ROS) of HCT 15 cells exposed to p -Coumaric acid was evaluated using 2', 7'-dichlorfluorescein- diacetate. Mitochondrial membrane potential of HCT-15 was assessed using rhodamine-123 with the help of flow cytometry. Lipid layer breaks associated with p -Coumaric acid treatment was quantified using the dye merocyanine 540. Apoptosis was confirmed and quantified using flow cytometric analysis of HCT 15 cells subjected to p -Coumaric acid treatment after staining with YO-PRO-1. RESULTS: Antiproliferative test showed p -Coumaric acid has an inhibitory effect on HCT 15 and HT 29 cells with an IC50 (concentration for 50% inhibition) value of 1400 and 1600 μmol/L respectively. Colony forming assay revealed the time-dependent inhibition of HCT 15 and HT 29 cells subjected to p -Coumaric acid treatment. Propidium iodide staining of treated HCT 15 cells showed increasing accumulation of apoptotic cells (37.45 ± 1.98 vs 1.07 ± 1.01) at sub-G1 phase of the cell cycle after p -Coumaric acid treatment. HCT-15 cells observed with photomicrograph and scanning electron microscope showed the signs of apoptosis like blebbing and shrinkage after p -Coumaric acid exposure. Evaluation of the lipid layer showed increasing lipid layer breaks was associated with the growth inhibition of p -Coumaric acid. A fall in mitochondrial membrane potential and increasing ROS generation was observed in the p -Coumaric acid treated cells. Further apoptosis evaluated by YO-PRO-1 staining also showed the timedependent increase of apoptotic cells after treatment. CONCLUSION: These results depicted that p -Coumaric acid inhibited the growth of colon cancer cells by inducing apoptosis through ROS-mitochondrial pathway
format Article
author Jaganathan, Saravana Kumar
Supriyanto, Eko
Mandal, Mahitosh
author_facet Jaganathan, Saravana Kumar
Supriyanto, Eko
Mandal, Mahitosh
author_sort Jaganathan, Saravana Kumar
title Events associated with apoptotic effect of p -Coumaric acid in HCT-15 colon cancer cells
title_short Events associated with apoptotic effect of p -Coumaric acid in HCT-15 colon cancer cells
title_full Events associated with apoptotic effect of p -Coumaric acid in HCT-15 colon cancer cells
title_fullStr Events associated with apoptotic effect of p -Coumaric acid in HCT-15 colon cancer cells
title_full_unstemmed Events associated with apoptotic effect of p -Coumaric acid in HCT-15 colon cancer cells
title_sort events associated with apoptotic effect of p -coumaric acid in hct-15 colon cancer cells
publisher Baishideng Publishing Group Co. Limited
publishDate 2013
url http://eprints.utm.my/id/eprint/50550/1/EkoSupriyanto2013_Eventsassociatedwithapoptotic.pdf
http://eprints.utm.my/id/eprint/50550/
http://dx.doi.org/10.3748/wjg.v19.i43.7726
_version_ 1643652807406911488
score 13.211869

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