Circular RNA_0025843 Alleviated Cigarette Smoke Extract Induced Bronchoalveolar Epithelial Cells Ferroptosis

Abstract

Purpose

Circular RNA (circRNA) plays an important role in various biological processes. However, their functions in cigarette smoke extract (CSE) induced human normal lung epithelial cells (BEAS-2B) injury remain vague. The study aimed to explore circRNA expression profiles and reveal their potential roles in CSE-treated BEAS-2B cells.

Methods

5% CSE exposure for 24 hours were used to build the BEAS-2B cells ferroptosis model. Differentially expressed circRNAs (DECs) were identified by next-generation RNA sequencing. Six randomly selected DECs were validated via quantitative reverse transcription polymerase chain reaction (qRT-PCR). Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis and Gene Ontology (GO) analysis were conducted to clarify the potential functions of the DECs. Furthermore, the role of hsa_circ_0025843 in CSE-related BEAS-2B cells ferroptosis was confirmed.

Results

5% CSE exposure induced BEAS-2B cells ferroptosis. Fifty-one up-regulated cirRNAs and 80 down-regulated circRNAs were revealed in CSE-treated BEAS-2B cells. Hsa_circ_0003461, hsa_circ_0007548, hsa_circ_0025843, hsa_circ_0068896, hsa_circ_0005832, and hsa_circ_0053378 were selected randomly to validate the reliability of next-generation RNA sequencing by qRT-PCR. After KEGG pathway analysis, DECs were found to participate in the process of EGFR tyrosine kinase inhibitor resistance and glycerophospholipid metabolism. The knockdown of hsa_circ_0025843 significantly alleviated CSE-induced BEAS-2B cells ferroptosis.

Conclusion

The study indicated the circRNA expression profiles in CSE-treated BEAS-2B cells. Hsa_circ_0025843 alleviated CSE induced BEAS-2B cells ferroptosis, which might be a potential therapeutic target of CSE related lung injury.

Keywords:

• cigarette smoke extract
• Circular RNA
• next-generation RNA sequencing
• lung injury
• ferroptosis

Abbreviations

BEAS-2B, human normal lung epithelial cells; CSE, cigarette smoke extract; circRNA, Circular RNA; PCR, polymerase chain reaction; GO, Gene Ontology; KEGG, Kyoto Encyclopedia of Genes and Genomes; ceRNA, competing endogenous RNA; BP, biological process; CC, cellular component; MF, molecular function; COPD, chronic obstructive pulmonary disease; ROS, reactive oxygen species; MMP, mitochondrial membrane potential; ACSL4, Acyl-CoA synthetase long-chain family member 4; GPX4, glutathione peroxidase 4.

Ethics Approval and Consent to Participate

This article does not contain any studies with human participants or animals performed by any of the authors.

Disclosure

The authors have no relevant financial or non-financial interests to disclose for this work.

Additional information

Funding

This study was funded by the National Natural Science Foundation of China (Grant number: 82170101) and the Joint Funds for the Innovation of Science and Technology, Fujian Province (Grant number: 2019Y9116).