Identifying crucial lncRNAs and mRNAs in hypoxia-induced A549 lung cancer cells and investigating their underlying mechanisms via high-throughput sequencing
Lin Lin, Lili Deng, Yongxia Bao
Abstract
Rapid proliferation and outgrowth of tumor cells frequently result in localized hypoxia, which has been implicated in the progression of lung cancer. The present study aimed to identify key long non-coding RNAs (lncRNAs) and messenger RNAs (mRNAs) involved in hypoxia-induced A549 lung cancer cells, and to investigate their potential underlying mechanisms of action.
Introduction
Lung cancer, a malignant tumor disease with a high degree of malignancy, has caused a huge health burden and economic pressure worldwide. Its mortality rate is up to 13.5%, making lung cancer one of the most fatal cancer types [1]. Despite significant advances in the research of lung cancer in the medical field, the causes, development mechanisms, and effective treatment methods of lung cancer still remain challenging. Despite significant and groundbreaking progress in the medical field of lung cancer treatment, unfortunately, the five-year survival rate for patients remains unfavorable [2].
Materials and method
Cell culture and hypoxia treatment
The human pulmonary adenocarcinoma cell line A549 was obtained from the cell bank of the Chinese Academy of Sciences. Cells are cultured in F-12K medium supplemented with 10% fetal bovine serum (FBS) and 1% Penicillin-Streptomycin (PS). Upon arrival, the cells are first allowed to rest overnight in the incubator to facilitate their adherence to the flask walls. The next day, the culture medium is aspirated, and 1 mL of 0.25% trypsin solution is added (sufficient to cover the entire bottom of the flask).
Results
The statistical power of this experimental design, calculated in RNASeqPower is 0.83. A flowchart for analyses is presented in Fig 1.
Discussion
Under hypoxic conditions, tumor cells activate the HIF signaling pathway, which not only accelerates tumor growth and enhances invasiveness but also promotes tumor metastasis. Despite our current understanding of the relationship between hypoxia and cancer, the molecular mechanisms underlying hypoxia-induced tumorigenesis remain incompletely understood. Exploring and refining these molecular mechanisms could provide novel therapeutic and preventive strategies for lung cancer. In this study, the differentially expressed lncRNAs and mRNAs in hypoxic-induced A549 lung cancer cells were screened by high-throughput sequencing, and the possible mechanism of action was explored. Finally, we identified 1155 mRNAs and 215 lncRNAs differentially expressed in hypoxia vs. control groups.
Acknowledgments
We thank all the participants and their family for their cooperation.
Citation: Lin L, Deng L, Bao Y (2024) Identifying crucial lncRNAs and mRNAs in hypoxia-induced A549 lung cancer cells and investigating their underlying mechanisms via high-throughput sequencing. PLoS ONE 19(9): e0307954. https://doi.org/10.1371/journal.pone.0307954
Editor: Qi Zhao, University of Science and Technology Liaoning, CHINA
Received: March 7, 2024; Accepted: July 1, 2024; Published: September 5, 2024
Copyright: © 2024 Lin et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Data Availability: The high-throughput sequencing data generated in this study are publicly accessible via the NCBI SRA database with the accession number SRP411127, accessible at the following link: https://dataview.ncbi.nlm.nih.gov/object/PRJNA906305?reviewer=dm5r2erlkr9hvdu6vpa4jtsolm.
Funding: We were fortunate to receive funding from the Natural Science Foundation of Heilongjiang Province of China (Grant No. LH2020H060). This funding supported sequencing services and partial experimental consumables for our research, but the funders had no involvement in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Competing interests: The authors have declared that no competing interests exist.
