Mechanism of hydrogen on cervical cancer suppression revealed by high‑throughput RNA sequencing

Chu,Jing; Gao,Jinghai; Wang,Jing; Li,Lingling; Chen,Guoqiang; Dang,Jianhong; Wang,Zhifeng; Jin,Zhijun; Liu,Xiaojun

doi:10.3892/or.2021.8092

Mechanism of hydrogen on cervical cancer suppression revealed by high‑throughput RNA sequencing

Authors:
- Jing Chu
- Jinghai Gao
- Jing Wang
- Lingling Li
- Guoqiang Chen
- Jianhong Dang
- Zhifeng Wang
- Zhijun Jin
- Xiaojun Liu
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Affiliations: Department of Obstetrics and Gynaecology, Changzheng Hospital, Naval Medical University, Shanghai 200003, P.R. China
Published online on: May 26, 2021 https://doi.org/10.3892/or.2021.8092
Article Number: 141
Copyright: © Chu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Cervical cancer is considered one of the diseases with the highest mortality among women and with limited treatment options. Hydrogen (H₂) inhalation has been reported to have a variety of tumor‑suppressive effects, but the exact mechanism remains unclear. In the present study, HeLa cervical cancer cells and HaCaT keratinocytes treated with H₂, and a HeLa xenograft mouse model subjected to H₂ inhalation were established. TUNEL, Cell Counting Kit‑8 and Ki67 staining assays were used to detect cell apoptosis and proliferation. Oxidative stress was determined according to the levels of reactive oxygen species, malondialdehyde and superoxide dismutase. Tumor growth was recorded every 3 days, and the excised tumors were stained with hematoxylin and eosin. High‑throughput RNA sequencing and subsequent Gene Ontology (GO) enrichment analysis were performed in HeLa‑treated and un‑treated HeLa cells. The expression of hypoxia‑inducible factor (HIF)‑1α and NF‑κB p65 was verified by western blotting, immunohistochemistry and reverse transcription‑quantitative PCR. The results revealed an increased apoptosis rate, and reduced cell proliferation and oxidative stress in H₂‑treated HeLa cells but not in HaCaT cells. Similarly, decreased tumor growth and cell proliferation, and enhanced cell apoptosis were observed in H2‑treated HeLa tumors. RNA sequencing and GO analysis suggest that downregulated HIF1A (HIF‑1α mRNA) and RelA (NF‑κB p65) levels, and reduced NF‑κB signaling were associated with the antitumor effect of H₂. Finally, decreased HIF‑1α and NF‑κB p65 expression both at the transcriptional and translational levels were observed in H₂‑treated HeLa cells and in HeLa‑derived tumors. In conclusion, the present study reveals a novel mechanism of H₂ against cervical cancer, which may serve as a potential therapeutic target in clinical practice.

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