Alkylation repair homolog 3-regulated esophageal squamous cell carcinoma associated long non-coding RNA 1 is required for maintaining the stemness of esophageal cancer

Yuanbo Cui¹, Yanan Lou², Pengju Lv¹, Wei Cao^1*

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¹ Translational Medicine Center, Zhengzhou Central Hospital Affiliated to Zhengzhou University, Zhengzhou 450007, China

² Department of Breast Surgery, Zhengzhou Central Hospital Affiliated to Zhengzhou University, Zhengzhou, 450007, China

GPD 2023, 2(1), 305 https://doi.org/10.36922/gpd.305

Submitted: 28 December 2022 | Accepted: 21 February 2023 | Published: 13 March 2023

© 2023 by the Author(s). This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution 4.0 International License ( https://creativecommons.org/licenses/by/4.0/ )

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Abstract

N1-methyladenosine (m1A) RNA modification represents one of the essential post-transcriptional modifications in gene expression regulation. Long non-coding RNAs (lncRNAs) are involved in the development of malignant tumors, including esophageal cancer (ESCA). However, whether m1A can regulate that lncRNA in cancer cells remains unclear. ESCA cell lines TE1 and KYSE70 were used for functional experiments. The mRNA and protein levels were detected by quantitative reverse transcription polymerase chain reaction and Western blot, respectively. Colony formation and tumor sphere formation assays were used for evaluating ESCA stemness. The m1A modification on esophageal squamous cell carcinoma associated long non-coding RNA 1 (ESCCAL-1) transcript was examined by methylated RNA immunoprecipitation. In this study, we report that RNA m1A demethylase alkylation repair homolog 3 (ALKBH3)-mediated ESCCAL-1 is implicated in maintaining stem cell-like properties of ESCA. Clinically, ESCCAL-1 was up-regulated in ESCA and positively correlated with tumor stage. In addition, patients with higher ESCCAL-1 expression in tumors had shorter median survival. Functionally, the knockdown of ESCCAL-1 attenuated the stemness of ESCA cells as indicated by decreased sphere formation and colony formation capacities, while overexpression of ESCCAL-1 elicits the opposite biological effects. Moreover, ESCCAL-1 manipulation positively regulated both mRNA and protein levels of KLF4 and CD44, two stemness-related markers. Mechanistically, ALKBH3 upregulated ESCCAL-1 expression in an m1A demethylation-dependent manner. Notably, the downregulation of ALKBH3 mimicked the effects of ESCCAL-1 deficiency on ESCA stemness, and this phenomenon is significantly reversed by the enforced expression of ESCCAL-1. Our results revealed the role of m1A-mediated ESCCAL-1 in ESCA self-renewal, which expands the understanding of lncRNA post-transcriptional modification in cancer development.

Keywords

ALKBH3

N1-methyladenosine

ESCCAL-1

Stemness

Esophageal cancer

Funding

Medical Science and Technology Project of Henan Province

Key Project of Higher Education in Henan Province

Conflict of interest

The authors declare that there are no competing interests.

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