Related Experiment Video
Updated: May 9, 2026
Detection of Total Reactive Oxygen Species in Adherent Cells by 2',7'-Dichlorodihydrofluorescein Diacetate Staining
Published on: June 23, 2020
Reactive oxygen species mediate tolfenamic acid-induced apoptosis in human colorectal cancer cells
Jin Boo Jeong1, Jieun Choi, Seung Joon Baek
1Department of Nutrition and Food Science, College of Agriculture and Natural Resources, University of Maryland, College Park, MD 20742, USA.
Abstract:
Several studies have shown substantial evidences that non-steroidal anti-inflammatory drugs (NSAIDs) exert anticancer effects by generating reactive oxygen species (ROS). Tolfenamic acid (TA) is one of the traditional NSAIDs widely used for treatment of migraine. TA has anti-cancer activities in several human cancer models. In this study, we report that generation of ROS by TA leads to apoptosis through modulation of several pathways in human colorectal cancer cells. TA induced rapid generation of intracellular ROS and led to an increase of phosphorylation of H2AX, a tail moment of comet and distribution of fragmented genomic DNA traces. Treatment of N-acetyl-l-cysteine (NAC) abolished TA-induced phosphorylation of H2AX and apoptosis. Treatment of TA resulted in an increase of nuclear factor-kappaB (NF-κB) transcriptional activity through inhibitor of kappa B (IκB-α) degradation and subsequent p65 nuclear translocation. In addition, TA increased apoptosis-inducing activating transcription factor 3 (ATF3) expression. However, the treatment of NAC abolished TA-mediated NF-κB activation and ATF3 expression and chemical inhibition of NF-κB or knockdown of p65 significantly attenuated TA-induced ATF3 expression. Our finding indicates that ROS-mediated DNA damage and subsequent activation of NF-κB and ATF3 expression plays a significant role in TA-induced apoptosis in human colorectal cancer cells.
Insights
Tolfenamic acid, a non-steroidal anti-inflammatory drug, triggers cancer cell death by generating reactive oxygen species (ROS). This process involves DNA damage and activation of key pathways, offering potential for colorectal cancer treatment.
Area of Science:
- Oncology
- Molecular Biology
- Pharmacology
Background:
- Non-steroidal anti-inflammatory drugs (NSAIDs) are known to possess anticancer properties.
- Reactive oxygen species (ROS) generation is a proposed mechanism for NSAID-induced anticancer effects.
- Tolfenamic acid (TA) is an NSAID used for migraine treatment with observed anti-cancer activity.
Purpose of the Study:
- To investigate the role of ROS in Tolfenamic acid-induced apoptosis in human colorectal cancer cells.
- To elucidate the molecular pathways modulated by TA in colorectal cancer.
Main Methods:
- Assessing intracellular ROS generation and DNA damage markers (H2AX phosphorylation, comet assay).
- Evaluating the impact of N-acetyl-l-cysteine (NAC) on TA-induced effects.
- Analyzing nuclear factor-kappaB (NF-κB) activation and apoptosis-inducing activating transcription factor 3 (ATF3) expression.
Main Results:
- TA induced rapid ROS generation, DNA damage, and apoptosis in colorectal cancer cells.
- N-acetyl-l-cysteine (NAC) counteracted TA-induced DNA damage and apoptosis.
- TA activated NF-κB signaling and increased ATF3 expression, which were abrogated by NAC or NF-κB inhibition.
Conclusions:
- ROS generation is a key mediator of Tolfenamic acid-induced apoptosis in human colorectal cancer.
- The mechanism involves ROS-mediated DNA damage, subsequent NF-κB activation, and ATF3 expression.
- TA demonstrates potential as a therapeutic agent for colorectal cancer via ROS-dependent pathways.
Related Concept Videos
01:17The Extrinsic Apoptotic Pathway
01:11Pharmacogenetics of Drug Targets: β₂-Adrenergic Receptors, Apo E, Thymidylate Synthase
01:25Bioactivation and Tissue Toxicity