Pioglitazone, a peroxisome proliferator‑activated receptor γ agonist, induces cell death and inhibits the proliferation of hypoxic HepG2 cells by promoting excessive production of reactive oxygen species

Huang,Guohao; Zhang,Mengfan; Wang,Manzhou; Xu,Wenze; Duan,Xuhua; Han,Xinwei; Ren,Jianzhuang

doi:10.3892/ol.2024.14294

Pioglitazone, a peroxisome proliferator‑activated receptor γ agonist, induces cell death and inhibits the proliferation of hypoxic HepG2 cells by promoting excessive production of reactive oxygen species

Authors:
- Guohao Huang
- Mengfan Zhang
- Manzhou Wang
- Wenze Xu
- Xuhua Duan
- Xinwei Han
- Jianzhuang Ren
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Affiliations: Department of Interventional Radiology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan 450052 P.R. China
Published online on: February 19, 2024 https://doi.org/10.3892/ol.2024.14294
Article Number: 160
Copyright: © Huang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Hypoxia is a hallmark of solid tumors. Hypoxic cancer cells adjust their metabolic characteristics to regulate the production of cellular reactive oxygen species (ROS) and facilitate ROS‑mediated metastasis. Peroxisome proliferator‑activated receptor γ (PPARγ) is a nuclear receptor that regulates the transcription of fatty acid metabolism‑related genes that have a key role in the survival and proliferation function of hypoxic cancer cells. In the present study, mRNA expression in HepG2 cells under chemically induced hypoxia was assessed. The protein expression levels of hypoxia‑inducible factor 1α (HIF‑1α) were measured using western blotting. Following treatment with the PPARγ agonist pioglitazone, cell viability was assessed using a Cell Counting Kit‑8 assay, whilst cell proliferation and death were determined using 5‑ethynyl‑2'‑deoxyuridine incorporation staining, and calcein‑acetoxymethyl ester and propidium iodide staining, respectively. Cellular ROS production was assessed using dihydroethidium staining. Cobalt chloride was used to induce hypoxia in HepG2 cells, which was evaluated using HIF‑1α expression. The results revealed that the mRNA expression of PPARγ, CD36, acetyl‑co‑enzyme A dehydrogenase (ACAD) medium chain (ACADM) and ACAD short‑chain (ACADS) was downregulated in hypoxic HepG2 cells. The PPARγ agonist pioglitazone decreased the cell viability of hypoxic HepG2 cells by inhibiting cell proliferation and inducing cell death. Following treatment with the PPARγ agonist pioglitazone, hypoxic HepG2 cells produced excessive ROS. ROS‑mediated cell death induced by the PPARγ agonist pioglitazone was rescued with the antioxidant N‑acetyl‑L‑cysteine. The downregulated mRNA expression of PPARγ, CD36, ACADM and ACADS was not reverted by a PPARγ agonist in hypoxic HepG2 cells. By contrast, the PPARγ agonist suppressed the mRNA expression of BCL2, which was upregulated in hypoxic HepG2 cells. In summary, the PPARγ agonist stimulated excessive ROS production to inhibit cell proliferation and increase the death of hypoxic HepG2 cells by decreasing BCL2 mRNA expression, suggesting a negative association between PPARγ and BCL2 in the regulation of ROS production in hypoxic HepG2 cells.

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