H19 recruited N^(6)-methyladenosine(m^(6)A)reader YTHDF1 to promote SCARB1 translation and facilitate angiogenesis in gastric cancer

作     者：Rumeng Bai Miaomiao Sun Yuanyuan Chen Shuaishuai Zhuo Guoxin Song Tianjun Wang Zhihong Zhang 

作者机构：Department of PathologyThe First Affiliated Hospital of Nanjing Medical UniversityNanjingJiangsu 210029China Department of PathologyWuxi Maternity and Child Health Hospital Affiliated to Nanjing Medical UniversityWuxiJiangsu 214002China Department of BiochemistryNanjing Medical UniversityNanjingJiangsu 211112China 

出 版 物：《Chinese Medical Journal》 (中华医学杂志（英文版）)

年 卷 期：2023年第136卷第14期

页      面：1719-1731页

核心收录：

学科分类：1002[医学-临床医学] 100214[医学-肿瘤学] 10[医学] 

基　　金：Department of Biochemistry and Molecular Biology, Nanjing medical university, Nanjing, Jiangsu Province Nanjing Xinjia Medical Technology Co. Ltd 

主　　题：Gastric cancer H19 long non-coding RNA Angiogenesis YTHDF1 protein,human SCARB1 protein,human 6-methyladenine 

摘      要：Background:Angiogenesis is described as a complex process in which new microvessels sprout from endothelial cells of existing vasculature.This study aimed to determine whether long non-coding RNA(lncRNA)H19 induced the angiogenesis of gastric cancer(GC)and its possible mechanism.Methods:Gene expression level was determined by quantitative real-time polymerase chain reaction and western blotting.Cell counting kit-8,transwell,5-Ethynyl-2′-deoxyuridine(EdU),colony formation assay,and human umbilical vein endothelial cells(HUVECs)angiogenesis assay as well as Matrigel plug assay were conducted to study the proliferation,migration,and angiogenesis of GC in vitro and in vivo.The binding protein of H19 was found by RNA pull-down and RNA Immunoprecipitation(RIP).High-throughput sequencing was performed and next Gene Ontology(GO)as well as Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis was conducted to analyze the genes that are under H19 regulation.Methylated RIP(me-RIP)assay was used to investigate the sites and abundance among target mRNA.The transcription factor acted as upstream of H19 was determined through chromatin immunoprecipitation(ChIP)and luciferase assay.Results:In this study,we found that hypoxia-induced factor(HIF)-1a could bind to the promoter region of H19,leading to H19 overexpression.High expression of H19 was correlated with angiogenesis in GC,and H19 knocking down could inhibit cell proliferation,migration and angiogenesis.Mechanistically,the oncogenic role of H19 was achieved by binding with the N^(6)-methyladenosine(m^(6)A)reader YTH domain-containing family protein 1(YTHDF1),which could recognize the m^(6)A site on the 3′-untransated regions(3′-UTR)of scavenger receptor class B member 1(SCARB1)mRNA,resulting in over-translation of SCARB1 and thus promoting the proliferation,migration,and angiogenesis of GC cells.Conclusion:HIF-1a induced overexpression of H19 via binding with the promoter of H19,and H19 promoted GC cells proliferation,migration and angiogenesis through YTHDF1/SCARB1,which might be a beneficial target for antiangiogenic therapy for GC.