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Abstract：
BACKGROUND: Multiple genetic and epigenetic regulatory mechanisms are crucial in the development and tumorigenesis process. Transcriptional regulation often involves intricate relationships and networks with post-transcriptional regulatory molecules, impacting the spatial and temporal expression of genes. However, the synergistic relationship between transcription factors and N6-methyladenosine (m6A) modification in regulating gene expression, as well as their influence on the mechanisms underlying the occurrence and progression of non-small cell lung cancer (NSCLC), requires further investigation. The present study aimed to investigate the synergistic relationship between transcription factors and m6A modification on NSCLC.
METHODS: The transcription factor NFIC and its potential genes was screened by analyzing publicly available datasets (ATAC-seq, DNase-seq, and RNA-seq). The association of NFIC and its potential target genes were validated through ChIP-qPCR and dual-luciferase reporter assays. Additionally, the roles of NFIC and its potential genes in NSCLC were detected in vitro and in vivo through silencing and overexpression assays.
RESULTS: Based on multi-omics data, the transcription factor NFIC was identified as a potential tumor suppressor of NSCLC. NFIC was significantly downregulated in both NSCLC tissues and cells, and when NFIC was overexpressed, the malignant phenotype and total m6A content of NSCLC cells was suppressed, while the PI3K/AKT pathway was inactivated. Additionally, we discovered that NFIC inhibits the expression of METTL3 by directly binding to its promoter region, and METTL3 regulates the expression of KAT2A, a histone acetyltransferase, by methylating the m6A site in the 3\'UTR of KAT2A mRNA in NSCLC cells. Intriguingly, NFIC was also found to negatively regulate the expression of KAT2A by directly binding to its promoter region.
CONCLUSIONS: Our findings demonstrated that NFIC suppresses the malignant phenotype of NSCLC cells by regulating gene expression at both the transcriptional and post-transcriptional levels. A deeper comprehension of the genetic and epigenetic regulatory mechanisms in tumorigenesis would be beneficial for the development of personalized treatment strategies.
METHODS: The transcription factor NFIC and its potential genes was screened by analyzing publicly available datasets (ATAC-seq, DNase-seq, and RNA-seq). The association of NFIC and its potential target genes were validated through ChIP-qPCR and dual-luciferase reporter assays. Additionally, the roles of NFIC and its potential genes in NSCLC were detected in vitro and in vivo through silencing and overexpression assays.
RESULTS: Based on multi-omics data, the transcription factor NFIC was identified as a potential tumor suppressor of NSCLC. NFIC was significantly downregulated in both NSCLC tissues and cells, and when NFIC was overexpressed, the malignant phenotype and total m6A content of NSCLC cells was suppressed, while the PI3K/AKT pathway was inactivated. Additionally, we discovered that NFIC inhibits the expression of METTL3 by directly binding to its promoter region, and METTL3 regulates the expression of KAT2A, a histone acetyltransferase, by methylating the m6A site in the 3\'UTR of KAT2A mRNA in NSCLC cells. Intriguingly, NFIC was also found to negatively regulate the expression of KAT2A by directly binding to its promoter region.
CONCLUSIONS: Our findings demonstrated that NFIC suppresses the malignant phenotype of NSCLC cells by regulating gene expression at both the transcriptional and post-transcriptional levels. A deeper comprehension of the genetic and epigenetic regulatory mechanisms in tumorigenesis would be beneficial for the development of personalized treatment strategies.
摘要：
背景：多种遗传和表观遗传调控机制在发育和肿瘤发生过程中至关重要。转录调控通常涉及与转录后调控分子的复杂关系和网络，影响基因的时空表达。然而,转录因子与N6-甲基腺苷（m6A）修饰在调节基因表达中的协同关系，以及它们对非小细胞肺癌（NSCLC）发生和进展的潜在机制的影响，需要进一步调查。本研究旨在探讨转录因子与m6A修饰在非小细胞肺癌中的协同作用。
方法：通过分析公开可用的数据集（ATAC-seq，DNase-seq,和RNA-seq)。通过ChIP-qPCR和双荧光素酶报告基因测定验证了NFIC及其潜在靶基因的关联。此外,通过沉默和过表达试验,在体外和体内检测NFIC及其潜在基因在NSCLC中的作用.
结果：基于多组学数据，转录因子NFIC被鉴定为NSCLC的潜在肿瘤抑制因子。NFIC在NSCLC组织和细胞中显著下调,当NFIC过度表达时，NSCLC细胞的恶性表型和总m6A含量被抑制，而PI3K/AKT途径失活。此外,我们发现NFIC通过直接结合其启动子区来抑制METTL3的表达,和METTL3调节KAT2A的表达，组蛋白乙酰转移酶，通过甲基化NSCLC细胞中KAT2AmRNA的3'UTR中的m6A位点。有趣的是，还发现NFIC通过直接结合其启动子区负调节KAT2A的表达。
结论：我们的研究结果表明，NFIC通过在转录和转录后水平调节基因表达来抑制NSCLC细胞的恶性表型。对肿瘤发生中的遗传和表观遗传调控机制的更深入理解将有利于制定个性化治疗策略。
方法：通过分析公开可用的数据集（ATAC-seq，DNase-seq,和RNA-seq)。通过ChIP-qPCR和双荧光素酶报告基因测定验证了NFIC及其潜在靶基因的关联。此外,通过沉默和过表达试验,在体外和体内检测NFIC及其潜在基因在NSCLC中的作用.
结果：基于多组学数据，转录因子NFIC被鉴定为NSCLC的潜在肿瘤抑制因子。NFIC在NSCLC组织和细胞中显著下调,当NFIC过度表达时，NSCLC细胞的恶性表型和总m6A含量被抑制，而PI3K/AKT途径失活。此外,我们发现NFIC通过直接结合其启动子区来抑制METTL3的表达,和METTL3调节KAT2A的表达，组蛋白乙酰转移酶，通过甲基化NSCLC细胞中KAT2AmRNA的3'UTR中的m6A位点。有趣的是，还发现NFIC通过直接结合其启动子区负调节KAT2A的表达。
结论：我们的研究结果表明，NFIC通过在转录和转录后水平调节基因表达来抑制NSCLC细胞的恶性表型。对肿瘤发生中的遗传和表观遗传调控机制的更深入理解将有利于制定个性化治疗策略。
