Luteolin induces mitochondrial apoptosis in HT29 cells by inhibiting the Nrf2/ARE signaling pathway

Yang,Huan; Liu,Bing‑Fang; Xie,Fu‑Jia; Yang,Wei‑Lin; Cao,Nong

doi:10.3892/etm.2020.8464

Luteolin induces mitochondrial apoptosis in HT29 cells by inhibiting the Nrf2/ARE signaling pathway

Authors:
- Huan Yang
- Bing‑Fang Liu
- Fu‑Jia Xie
- Wei‑Lin Yang
- Nong Cao
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Affiliations: Department of General Surgery, First Hospital of Lanzhou University, Lanzhou, Gansu 730000, P.R. China, Department of Nuclear Magnetic Resonance, The Second Clinical Medical College of Lanzhou University, Lanzhou, Gansu 730000, P.R. China, Department of General Surgery, First Affiliated Hospital of Kunming Medical University, Kunming, Yunnan 650000, P.R China
Published online on: January 22, 2020 https://doi.org/10.3892/etm.2020.8464
Pages: 2179-2187
Copyright: © Yang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The aim of the current study was to investigate luteolin‑induced apoptosis and the molecular mechanisms underlying it in HT29 cells. A 3‑(4,5‑dimethylthiazol‑2‑yl)‑2,5‑diphenyltetrazolium bromide assay was used to assess the cytotoxicity of luteolin on HT29 cells, and a dichloro‑dihydro‑fluorescein diacetate assay was used to measure cellular levels of reactive oxygen species (ROS). The effects of luteolin on the mitochondrial membrane potential were also evaluated. Bax and Bcl‑2 mRNA expression were determined using reverse transcription‑quantitative PCR. Additionally, western blot analysis was performed to assess changes in cytochrome c and caspase‑3 protein expression. Localization of nuclear factor erythroid 2‑related factor 2 (Nrf2) in the nucleus was also assessed using immunofluorescence. Luteolin exhibited cytotoxicity on HT29 cells in a time‑ and concentration‑dependent manner. Additionally, ROS production was indicated to be increased and ROS scavenging was decreased, which resulted in a significant increase in the levels of ROS in the cells. The mitochondrial membrane potential was indicated to decrease following luteolin treatment. At the molecular level, luteolin significantly increased the mRNA expression of Bax and the protein expression of cytochrome c, caspase‑3, p47phox and p22phox. The results revealed that luteolin decreased Bcl‑2 protein expression and inhibited the nuclear localization of Nrf2. In conclusion, the current study indicated that luteolin inhibited HT29 cell proliferation and induced apoptosis via the mitochondrial pathway.

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