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ALKBH5 promotes the progression of infantile hemangioma through regulating the NEAT1/miR-378b/FOSL1 axis

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Abstract

Our work aims to investigate long non-coding RNA (lncRNA) N6-methyladenosine (m6A) modification and its role in infantile hemangioma (IH). The mRNA and protein expression levels were assessed using quantitative real-time polymerase chain reaction, western blot and immunohistochemistry. Me-RIP assay was performed to evaluate lncRNA NEAT1 m6A levels. Cell proliferation, migration and invasion were evaluated using cell counting kit-8 assay, transwell migration and invasion assay, respectively. Photo-activatable ribonucleoside-enhanced crosslinking and immunoprecipitation assay was conducted to verify the binding relationship between lncRNA nuclear paraspeckle assembly transcript 1 (NEAT1) and ALKBH5 (an RNA demethylase). The binding relationship between lncRNA NEAT1, microRNA (miR)-378b and FOS-like antigen 1 (FOSL1) was verified using dual-luciferase reporter gene assay and/or RNA immunoprecipitation assay. ALKBH5, lncRNA NEAT1 and FOLS1 expression was elevated in IH tissues, while miR-378b was downregulated. ALKBH5 knockdown suppressed cell proliferation, migration and invasion of IH cells, while promoting cell apoptosis. ALKBH5 promoted lncRNA NEAT1 expression by reducing the m6A modification of lncRNA NEAT1. In addition, miR‐378b was the target of lncRNA NEAT1, and its overexpression reversed the promotion effect of lncRNA NEAT1 overexpression on IH cell tumor-like behaviors. Moreover, FOLS1 was the target of miR-378b, and its overexpression reversed the inhibitory effect of miR-378b overexpression on IH cell tumor-like behaviors in vitro. ALKBH5 might have great potential as therapeutic target for IH, since ALKBH5 silencing suppressed IH progression by regulation of the NEAT1/miR-378b/FOSL1 axis.

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Data availability

All data generated or analyzed during this study are included in this article. The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

Code availability

Not applicable.

Material availability

All data generated or analyzed during this study are included in this article. The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

Abbreviations

ALKBH5:

Alkylation repair homolog protein 5

ANOVA:

Analysis of variance

CCK8:

Cell counting kit-8

ceRNAs:

Competing endogenous RNAs

DMEM:

Dulbecco's modified eagle media

FOSL1:

FOS-like antigen 1

FAR1:

Fos-related antigen 1

IHC:

Immunohistochemistry

IH:

Infantile hemangioma

lncRNA:

Long non-coding RNA

Me-RIP:

Methylated RNA binding protein immunoprecipitation

miR:

MicroRNA

NF1:

Neurofibromatosis 1

ncRNA:

Non-coding RNA

m6A:

N6-Methyladenosine

NEAT1:

Nuclear paraspeckle assembly transcript 1

PAR-CLIP:

Photo-activatable ribonucleoside-enhanced crosslinking and immunoprecipitation

qRT-PCR:

Quantitative real-time polymerase chain reaction

RIP:

RNA immunoprecipitation

SDS-PAGE:

Sodium dodecyl sulfate–polyacrylamide gel electrophoresis

SD:

Standard deviation

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Acknowledgements

We would like to give our sincere gratitude to the reviewers for their constructive comments.

Funding

This work was supported by Scientific Research Project of Hunan Provincial Health Commission (No.202206023142).

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Authors and Affiliations

  1. Department of Fetal and Neonatal Surgery, Hunan Children’s Hospital, No.86, Ziyuan Road, Yuhua District, Changsha, 410007, Hunan, People’s Republic of China

    Kun Peng, Ren-Peng Xia, Fan Zhao, Yong Xiao, Ti-Dong Ma, Ming Li, Yong Feng & Chong-Gao Zhou

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  1. Kun Peng
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  3. Fan Zhao
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  8. Chong-Gao Zhou
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Contributions

KP: Conceptualization; Writing-original draft; Methodology; Formal analysis; RPX: Supervision; Validation; FZ: Data curation; YX: Resources; TDM: Investigation; ML: Software; YF: Visualization; CGZ: Project administration; Funding acquisition; Writing-review and editing. All authors have read and approved the final version of this manuscript to be published.

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Correspondence to Chong-Gao Zhou.

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This study passed the review of the Ethics Committee of Hunan children's Hospital before enrollment of patients and all participants signed informed consent. The animal studies were approved by Ethics Committee of Hunan children's Hospital.

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Peng, K., Xia, RP., Zhao, F. et al. ALKBH5 promotes the progression of infantile hemangioma through regulating the NEAT1/miR-378b/FOSL1 axis. Mol Cell Biochem 477, 1527–1540 (2022). https://doi.org/10.1007/s11010-022-04388-2

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