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Gallic acid induced apoptotic events in HCT-15 colon cancer cells

Subramanian, A. P. and Jaganathan, S. K. and Mandal, M. and Supriyanto, E. and Muhamad, I. I. (2016) Gallic acid induced apoptotic events in HCT-15 colon cancer cells. World Journal of Gastroenterology, 22 (15). pp. 3952-3961. ISSN 1007-9327

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Abstract

AIM: To investigate the inhibitory action of diet-derived phenolic compound gallic acid (GA) against HCT-15 colon cancer cells. METHODS: The antiproliferative effect of GA against colon cancer cells was determined by performing thiazolyl blue tetrazolium bromide (MTT) assay. The colony forming ability of GA treated colon cancer cells was evaluated using the colony forming assay. The cell cycle changes induced by GA in HCT-15 cells were analyzed by propidium iodide staining. Levels of reactive oxygen species (ROS) and mitochondrial membrane potential of HCT-15 exposed to GA was assessed using 2',7'-dichlorfluorescein-diacetate and rhodamine-123 respectively, with the help of flow cytometry. Morphological changes caused by GA treatment in the colon cancer cells were identified by scanning electron microscope and photomicrograph examination. Apoptosis was confirmed using flow cytometric analysis of GA treated HCT-15 cells after staining with Yo-Pro-1. RESULTS: MTT assay results illustrated that GA has an inhibitory effect on HCT-15 cells with IC50 value of 740 μmol/L. A time-dependent inhibition of colony formation was evident with GA treatment. Cell cycle arrest was evident from the accumulation of GA treated HCT-15 cells at sub-G1 phase (0.98 ± 1.03 vs 58.01 ± 2.05) with increasing exposure time. Flow cytometric analysis of GA treated HCT-15 cells depicted early events associated with apoptosis like lipid layer breakage and fall in mitochondrial membrane potential apart from an increase in the generation of ROS which were in a time dependent manner. SEM and photomicrograph images of the GA-treated cells displayed membrane blebbing and cell shrinking characteristics of apoptosis. Further apoptosis confirmation by Yo-Pro-1 staining also showed the time-dependent increase of apoptotic cells after treatment. CONCLUSION: These results show that GA induced ROS dependent apoptosis and inhibited the growth of colon cancer cells.

Item Type:Article
Uncontrolled Keywords:2,7 dichlorfluorescein diacetate, acetoacetic acid, gallic acid, propidium iodide, reactive oxygen metabolite, rhodamine 123, unclassified drug, antineoplastic agent, gallic acid, reactive oxygen metabolite, antiproliferative activity, apoptosis, Article, cancer cell, cell count, cell cycle arrest, cell cycle G1 phase, cell cycle G2 phase, cell proliferation, cell viability, colony forming unit, colorectal carcinoma, controlled study, flow cytometry, human, human cell, IC50, microphotography, mitochondrial membrane potential, MTT assay, scanning electron microscopy, time-dependent inhibition, adenocarcinoma, apoptosis, cell survival, Colonic Neoplasms, dose response, drug effects, G1 phase cell cycle checkpoint, metabolism, oxidative stress, time factor, tumor cell line, ultrastructure, Adenocarcinoma, Antineoplastic Agents, Apoptosis, Cell Line, Tumor, Cell Proliferation, Cell Survival, Colonic Neoplasms, Dose-Response Relationship, Drug, G1 Phase Cell Cycle Checkpoints, Gallic Acid, Humans, Inhibitory Concentration 50, Membrane Potential, Mitochondrial, Oxidative Stress, Reactive Oxygen Species, Time Factors
Subjects:Q Science > QH Natural history > QH301 Biology
Divisions:Biosciences and Medical Engineering
ID Code:72618
Deposited By: Haliza Zainal
Deposited On:27 Nov 2017 01:40
Last Modified:27 Nov 2017 01:40

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