Gallic acid induced apoptotic events in HCT-15 colon cancer cells
Aruna Priyadharshni Subramanian1, Saravana Kumar Jaganathan1, Mahitosh Mandal1
1Aruna Priyadharshni Subramanian, Saravana Kumar Jaganathan, Eko Supriyanto, Ida Idayu Muhamad, IJN-UTM Cardiovascular Engineering Centre, Faculty of Biosciences and Medical Engineering, Universiti Teknologi Malaysia, Johor Bahru 81310, Malaysia.
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.
Insights
Gallic acid (GA) effectively inhibits HCT-15 colon cancer cell growth by inducing apoptosis. This study demonstrates GA's potential as an anti-cancer agent by triggering reactive oxygen species (ROS) production and cell cycle arrest.
Area of Science:
- Oncology
- Molecular Biology
- Natural Product Chemistry
Background:
- Diet-derived phenolic compounds are increasingly recognized for their therapeutic potential.
- Gallic acid (GA), a common plant phenolic, has shown various biological activities.
- The specific anti-cancer mechanisms of GA against colon cancer cells require further elucidation.
Purpose of the Study:
- To investigate the inhibitory action of gallic acid (GA) against HCT-15 colon cancer cells.
- To determine the underlying mechanisms of GA-induced anti-cancer effects.
- To evaluate GA's impact on cell proliferation, cell cycle, and apoptosis.
Main Methods:
- Antiproliferative effects were assessed using MTT and colony formation assays.
- Cell cycle distribution was analyzed via propidium iodide staining and flow cytometry.
- Reactive oxygen species (ROS) levels and mitochondrial membrane potential were measured using flow cytometry.
- Morphological changes and apoptosis were confirmed by scanning electron microscopy and Yo-Pro-1 staining.
Main Results:
- Gallic acid exhibited an inhibitory effect on HCT-15 cells with an IC50 of 740 μmol/L.
- GA treatment led to a time-dependent inhibition of colony formation and cell cycle arrest at the sub-G1 phase.
- Increased ROS generation, decreased mitochondrial membrane potential, and characteristic apoptotic morphological changes were observed.
- Apoptosis induction was confirmed by Yo-Pro-1 staining, showing a time-dependent increase in apoptotic cells.
Conclusions:
- Gallic acid effectively inhibits HCT-15 colon cancer cell growth.
- GA induces apoptosis in colon cancer cells in a ROS-dependent manner.
- These findings highlight GA's potential as a therapeutic agent for colon cancer.
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