Variant rs8400 enhances ALKBH5 expression through disrupting miR-186 binding and promotes neuroblastoma progression

作者：Qian Guan, Huiran Lin, Wenfeng Hua, Lei Lin, Jiabin Liu, Linqing Deng, Jiao Zhang, Jiwen Cheng, Zhonghua Yang, Yong Li, Jun Bian, Haixia Zhou, Suhong Li, Li Li, Lei Miao, Huimin Xia, Jing He, Zhenjian Zhuo · 发表于：Chinese Journal of Cancer Research · 年份：2023 · DOI：10.21147/j.issn.1000-9604.2023.02.05 · 被引用次数：52 · 研究领域：Neuroblastoma Research and Treatments、RNA modifications and cancer、Cancer, Hypoxia, and Metabolism

Objective AlkB homolog 5 (ALKBH5) has been proven to be closely related to tumors. However, the role and molecular mechanism of ALKBH5 in neuroblastomas have rarely been reported. Methods The potential functional single-nucleotide polymorphisms (SNPs) in ALKBH5 were identified by National Center for Biotechnology Information (NCBI) dbSNP screening and SNPinfo software. TaqMan probes were used for genotyping. A multiple logistic regression model was used to evaluate the effects of different SNP loci on the risk of neuroblastoma. The expression of ALKBH5 in neuroblastoma was evaluated by Western blotting and immunohistochemistry (IHC). Cell counting kit-8 (CCK-8), plate colony formation and 5-ethynyl-2’-deoxyuridine (EdU) incorporation assays were used to evaluate cell proliferation. Wound healing and Transwell assays were used to compare cell migration and invasion. Thermodynamic modelling was performed to predict the ability of miRNAs to bind to ALKBH5 with the rs8400 G/A polymorphism. RNA sequencing, N6-methyladenosine (m 6 A) sequencing, m 6 A methylated RNA immunoprecipitation (MeRIP) and a luciferase assay were used to identify the targeting effect of ALKBH5 on SPP1. Results ALKBH5 was highly expressed in neuroblastoma. Knocking down ALKBH5 inhibited the proliferation, migration and invasion of cancer cells. miR-186-3p negatively regulates the expression of ALKBH5, and this ability is affected by the rs8400 polymorphism. When the G nucleotide was mutated to A, the...