Jundishapur Journal of Medical Sciences

Jundishapur Journal of Medical Sciences

The Effect of High-Dose Ascorbic Acid on Necroptosis Pathway Activation in the HT29 Colon Cancer Cell Line

Document Type : Original Article

Authors
Dian Dayer 1
Zahra Shokati Eshkiki 2
Kosar Abbasi Samie 2
1 Cellular and Molecular Research Center,, Medical Basic Sciences Institute, Ahvaz Jundishapur University of Medical Sciences.
2 Alimentary Tract Research Center, Clinical Sciences Research Institute, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran.
10.22118/jsmj.2026.544167.3978
Abstract
Objective: The effect of high dosages of ascorbic acid on the expression of major necroptosis pathway genes in HT-29 colon cancer cells was studied. Materials and methods: HT-29 cells were cultured in DMEM-HG medium containing 10% FBS and 1% Pen/Strep and incubated at 37°C with 5% CO2. After reaching 90% density, the cells were trypsinised and cultured in triplicate in 12-well culture plates at 30,000 cells per well. The cells were divided into two groups: a control group that received only basal culture medium and a test group that received 5 mM ascorbic acid for 3 hours. After 24 hours, the cell supernatant and the cell pellet were collected for further analysis. LDH activity was assessed using the kinetic technique, and the proportion of apoptosis and necrosis was calculated using flow cytometry. Gene expression analysis was performed using real-time PCR. Statistical analysis was performed using GraphPad Prism software version 8 and a t-test. Results: Microscopic images and flow cytometry results confirmed the induction of necroptosis by ascorbic acid. In addition, ascorbic acid treatment significantly increased key necroptosis gene expression and LDH activity. Conclusion: High dosages of ascorbic acid activate necroptosis in HT-29 cells.
Keywords
Ascorbic acid
necroptosis
colon cancer
HT29 cell line
Subjects
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  • Receive Date 31 August 2025
  • Revise Date 10 December 2025
  • Accept Date 10 February 2026