Integrative analyses of N6-methyladenosine-associated single-nucleotide polymorphisms (m6A-SNPs) identify tumor suppressor gene AK9 in lung cancer
Integrative analyses of N6-methyladenosine-associated single-nucleotide polymorphisms (m6A-SNPs) identify tumor suppressor gene AK9 in lung cancer
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DOI：
10.1002/mc.23669
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发表时间：
2023-12-05
影响因子：
4.6
通讯作者：
Ma，Hongxia
中科院分区：
文献类型：
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作者：
Hua，Tingting;Zhang，Chang;Ma，Hongxia
N6-甲基腺苷(M6A)修饰被认为是人类肿瘤发生发展过程中最重要的表观遗传调控机制之一。然而，m6A相关单核苷酸多态(m6A-SNPs)与肺癌风险之间的关联在很大程度上仍不清楚。在这里，我们在13,793例肺癌患者和14,027名对照中识别了m6A-SNPs，并研究了这些m6A-SNPs与肺癌风险的关系。在电子计算机中，使用功能注释来识别因果M6A-SNPs和靶基因。采用甲基化RNA免疫沉淀法和实时定量聚合酶链式反应(MERIP-qPCR)检测不同SNP基因的m6A基因修饰水平。体外实验验证了靶基因在肺癌中的潜在作用。共检测到8794个m6A-SNPs，其中有9个易感基因座的397个SNPs与肺癌相关，其中包括6个新发现的基因座。生物信息学分析表明，6q21中的rs1321328位于AK9的m6A修饰位点附近，显著降低了AK9的表达(β= −为0.15，p= 2.78 × 10−8)。此外，肺癌组织中AK9的表达显著低于癌基因组图谱和南京肺癌队列中癌旁正常组织中的表达。MERIP-qPCR分析表明，与G等位基因相比，rs1321328的C等位基因可显著降低AK9的m6A修饰水平。体外实验证实了AK9在肺癌中的抑瘤作用。这些发现揭示了肺癌易感基因与m6A基因修饰相关的致病机制。
N6‐methyladenosine (m6A) modification has been identified as one of the most important epigenetic regulation mechanisms in the development of human cancers. However, the association between m6A‐associated single‐nucleotide polymorphisms (m6A‐SNPs) and lung cancer risk remains largely unknown. Here, we identified m6A‐SNPs and examined the association of these m6A‐SNPs with lung cancer risk in 13,793 lung cancer cases and 14,027 controls. In silico functional annotation was used to identify causal m6A‐SNPs and target genes. Furthermore, methylated RNA immunoprecipitation and quantitative real‐time polymerase chain reaction (MeRIP‐qPCR) assay was performed to assess the m6A modification level of different genotypes of the causal SNP. In vitro assays were performed to validate the potential role of the target gene in lung cancer. A total of 8794 m6A‐SNPs were detected, among which 397 SNPs in nine susceptibility loci were associated with lung cancer risk, including six novel loci. Bioinformatics analyses indicated that rs1321328 in 6q21 was located around the m6A modification site ofAK9and significantly reducedAK9expression (β= −0.15,p= 2.78 × 10−8). Moreover,AK9was significantly downregulated in lung cancer tissues than that in adjacent normal tissues of samples from the Cancer Genome Atlas and Nanjing Lung Cancer Cohort. MeRIP‐qPCR assay suggested that C allele of rs1321328 could significantly decrease the m6A modification level ofAK9compared with G allele. In vitro assays verified the tumor‐suppressing role ofAK9in lung cancer. These findings shed light on the pathogenic mechanism of lung cancer susceptibility loci linked with m6A modification.