Hesperetin induces apoptosis in lung squamous carcinoma cells via G<sub>2</sub>/M cycle arrest, inhibition of the Notch1 pathway and activation of endoplasmic reticulum stress

Xie,Qianlong; He,Ziming; Tan,Lingfang; Li,Min; Zhuang,Min; Liu,Chen; Chen,Sunhui; Jin,Long; Sui,Yuxia

doi:10.3892/ijmm.2025.5518

Hesperetin induces apoptosis in lung squamous carcinoma cells via G₂/M cycle arrest, inhibition of the Notch1 pathway and activation of endoplasmic reticulum stress

Authors:
- Qianlong Xie
- Ziming He
- Lingfang Tan
- Min Li
- Min Zhuang
- Chen Liu
- Sunhui Chen
- Long Jin
- Yuxia Sui
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Affiliations: Department of Pharmacy, Wuping County Hospital, Longyan, Fujian 363400, P.R. China, School of Pharmacy, Fujian Medical University, Fuzhou, Fujian 350122, China, Department of Clinical Laboratory, Fuzhou University Affiliated Provincial Hospital, Fuzhou, Fujian 350001, P.R. China, School of Basic Medical Sciences, Fujian Medical University, Fuzhou, Fujian 350122, P.R. China, Department of Pharmacy, Fuzhou University Affiliated Provincial Hospital, Fuzhou, Fujian 350001, P.R. China, Department of Pathology, Fuzhou University Affiliated Provincial Hospital, Fuzhou, Fujian 350001, P.R. China
Published online on: March 12, 2025 https://doi.org/10.3892/ijmm.2025.5518
Article Number: 77
Copyright: © Xie et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Hesperetin (HST), a natural flavonoid, has potent antitumor effects on lung adenocarcinoma; however, its effects on lung squamous cell carcinoma (LUSC) are currently unknown. The present study aimed to investigate the anticancer effects of HST on LUSC cells. The influence of 37.5, 75 and 150 µM HST on the H1703 cell line, and of 75, 150 and 300 µM HST on the H226 cell line was determined using the Cell Counting Kit‑8 method, cell cycle assay, JC‑1 mitochondrial membrane potential assay and Annexin V‑FITC/PI staining. DMSO‑treated cells were used as the control group. Western blotting was performed to detect the protein expression levels of cyclin B1, CDK1, Bcl‑2, Bax, caspase‑3, cleaved caspase‑3, phosphorylated‑eIF2α, eIF2α, glucose‑regulated protein 78, CHOP, Notch1 and Hes‑1. The relationship between endoplasmic reticulum stress (ERS), Notch1 signaling and apoptosis was examined using the ERS‑inhibitor 4‑phenylbutyric acid (4‑PBA; 500 µM) and the Notch1 signaling activator Jagged‑1 (4 µM). In vivo, mice were divided into control, HST (30, 60 and 90 mg/kg/q2d) and cisplatin (2 mg/kg/q2d) groups to evaluate the anti‑LUSC effects of HST. The results revealed that HST inhibited the viability of H226 and H1703 cells, leading to cell cycle arrest at the G₂/M phase and the induction of cell apoptosis. In addition, HST downregulated the Notch1 signaling pathway and increased ERS. In H1703 cells, 4‑PBA and Jagged‑1 reduced the expression of apoptosis‑related proteins, and Jagged‑1 also reduced the expression of ERS‑related proteins. In vivo, HST reduced tumor growth without any apparent toxic side effects. In conclusion, HST may exert its antitumor effects by inducing G₂/M cell cycle arrest and inhibiting the Notch1 signaling pathway to activate ERS‑induced apoptosis, making it a promising agent for treating LUSC.

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