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ALKBH5 decreases SLC7A11 expression by erasing m6A modification and promotes the ferroptosis of colorectal cancer cells

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Abstract

Background

Colorectal cancer (CRC) is the major subtype of gastrointestinal malignancy and involves cancer-related genes and signaling pathways to regulate ferroptosis. The present study was conducted to analyze the role of alkB homolog 5 (ALKBH5) in the ferroptosis of CRC cells and provide novel targets for CRC treatment.

Methods

The transcriptional and protein levels of ALKBH5 and solute carrier family 7 members 11 (SLC7A11) in tissues and cells were determined by qRT-PCR and Western blot assay. HCT116 and SW620 cells were transfected with ALKBH5 overexpression vectors to determine cell viability and levels of reactive oxygen species (ROS), Fe+, glutathione, and glutathione peroxidase 4 using cell counting kit-8, colony formation, fluorescence probe, assay kits, and Western blot assay. The N6-methyladenosine (m6A) level and the enrichment of m6A on SLC7A11 mRNA were measured by m6A quantitative analysis and m6A methylated RNA immunoprecipitation-qPCR, and the mRNA stability was determined after actinomycin D treatment. CRC cells were treated with the combination of SLC7A11 and ALKBH5 overexpression vectors to confirm the mechanism. Nude mice were subcutaneously injected with CRC cells overexpressing ALKBH5.

Results

ALKBH5 was downregulated in CRC and ALKBH5 overexpression promoted ROS release and ferroptosis. ALKBH5 erased the m6A modification on SLC7A11 mRNA to reduce the mRNA stability of SLC7A11, further reducing SLC7A11 expression. SLC7A11 overexpression reversed the promotive role of ALKBH5 overexpression in ferroptosis. ALKBH5 upregulation mitigated tumor growth in vivo.

Conclusions

ALKBH5 reduced SLC7A11 transcription by erasing m6A modification, thus promoting the ferroptosis of CRC cells.

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Data availability

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

Abbreviations

CRC:

Colorectal cancer

PCD:

Programmed cell death

ROS:

Reactive oxygen species

ALKBH5 :

AlkB homolog 5

SLC7A11 :

Solute carrier family 7 member 11

GSH:

Glutathione

GPX4:

Glutathione peroxidase 4

RPMI:

Roswell Park Memorial Institute

CCK-8:

Cell counting kit-8

DCFH-DA:

2’-7’ Dichlorofluorescein diacetate

qRT-PCR:

Quantitative real-time polymerase chain reaction

Ct:

Cycle threshold

MeRIP:

Methylated RNA immunoprecipitation

IHC:

Immunohistochemistry

COAD:

Colon adenocarcinoma

ANOVA:

Analysis of variance

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Authors and Affiliations

  1. Department of Oncology, Minhang Branch, Zhongshan Hospital, Fudan University, No.170 Xinsong Road, Minhang District, Shanghai, 201100, China

    Jing Luo, Hongmei Yu, Zhen Yuan, Tao Ye & Bo Hu

Authors
  1. Jing Luo
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  2. Hongmei Yu
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  3. Zhen Yuan
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  4. Tao Ye
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  5. Bo Hu
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Contributions

JL, HY and ZY designed and performed the research; JL and HY analyzed the data; JL, TY and BH manipulated the figures; JL wrote the manuscript. All authors reviewed and approved the final manuscript.

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Correspondence to Tao Ye or Bo Hu.

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Conflict of interest

All the authors declare that they have no competing interests.

Ethical approval and consent to participate

All animal experiments were conducted following the requirements from Guidelines for the care and use of laboratory animals in biomedical research [23] and got the ratification by the Animal Ethics Committee of Minhang Hospital.

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Luo, J., Yu, H., Yuan, Z. et al. ALKBH5 decreases SLC7A11 expression by erasing m6A modification and promotes the ferroptosis of colorectal cancer cells. Clin Transl Oncol 25, 2265–2276 (2023). https://doi.org/10.1007/s12094-023-03116-6

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  • DOI: https://doi.org/10.1007/s12094-023-03116-6

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