Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2025 Aug 12:S2090-1232(25)00599-5.
doi: 10.1016/j.jare.2025.08.010. Online ahead of print.

ALKBH5 modulates COL5A1 mRNA stability via m6A demethylation to promote tobacco carcinogen NNK-induced lung cancer occurrence and progression

Yiheng Du  1 , Lirong Tu  1 , Jiaxin Zhou  1 , Huanliang Huang  2 , Enwu Xu  3 , Dexiong Chen  2 , Xin Li  4 , Qiaoyuan Yang  5
Affiliations

ALKBH5 modulates COL5A1 mRNA stability via m6A demethylation to promote tobacco carcinogen NNK-induced lung cancer occurrence and progression

Yiheng Du et al. J Adv Res. .

Abstract

Introduction: The tobacco-specific carcinogen 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), has been extensively studied for its carcinogenic role in lung cancer. However, the involvement of N6-methyladenosine (m6A) methylation in NNK-induced carcinogenesis remains unclear.

Objectives: The aim of this study is to investigatewhether NNK promotes lung carcinogenesis by dysregulating m6A methylation, focusing on the demethylase ALKBH5 and its target COL5A1.

Methods: Levels of m6A were analyzed in NNK-exposed (100 mg/L, 24 h) Beas-2B bronchial epithelial cells, NNK-induced malignant-transformed Beas-2B (2B-NNK) cells, and paired lung tumor tissues. Methylated RNA immunoprecipitation sequencing, RNA sequencing, and bioinformatics identified COL5A1 as a key target. We performed proliferation, apoptosis, migration and invasion experiments involving COL5A1 and ALKBH5 knockdown and COL5A1 stability experiments. We then overexpressed COL5A1 and knocked down ALKBH5 for phenotypic rescue experiments.

Results: Levels of m6A were significantly reduced in NNK-exposed Beas-2B cells, 2B-NNK cells, and human lung tumor tissues. COL5A1 exhibited hypomethylation and overexpression in 2B-NNK cells. COL5A1 knockdown suppressed 2B-NNK cell proliferation, migration, and invasion, while increasing apoptosis. The m6A demethylase ALKBH5 was upregulated in 2B-NNK cells. Its knockdown elevated global m6A levels, reduced COL5A1 mRNA stability, and reversed malignant phenotypes. COL5A1 overexpression also enhanced the proliferation, migration, and invasion abilities of 2B-NNK cells, whereas concurrent downregulation of ALKBH5 expression resulted in the restoration of these enhancements.

Conclusion: ALKBH5 drives NNK-induced lung cancer via m6A demethylation of COL5A1, promoting tumor progression. Targeting the ALKBH5-COL5A1 axis may offer therapeutic strategies against tobacco-related cancers.

Keywords: ALKBH5; COL5A1; Lung cancer; NNK; m6A modification.

PubMed Disclaimer

Conflict of interest statement

Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.