PO.CL12.03 · 临床研究
靶向致癌性RNA m6A阅读器YTHDF2通过阻止p53降解抑制口腔鳞状细胞癌
Targeting the oncogenic RNA m6A reader YTHDF2 inhibits oral squamous cell carcinoma by preventing p53 degradation
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
背景：N6-甲基腺苷（m6A）RNA修饰机制已成为癌症生物学中的关键调节因子，然而，m6A"阅读器"蛋白在口腔鳞状细胞癌（OSCC）中的功能尚未完全阐明。YTH结构域家族蛋白2（YTHDF2）是一种重要的m6A结合蛋白（m6A"阅读器"），它促进甲基化mRNA的降解，并通过靶向使一组肿瘤抑制mRNA不稳定而与肿瘤发生相关。然而，YTHDF2如何促进OSCC增殖并调控p53活性的确切机制亟待阐明。本研究探讨了YTHDF2在OSCC中的致癌作用，并评估其作为诊断生物标志物和治疗靶点的潜力。
方法：通过定量实时PCR（qRT-PCR）和Western blot检测OSCC组织、配对的邻近非癌标本以及OSCC细胞系中YTHDF2的mRNA和蛋白水平。此外，采用TCGA-OSCC数据集评估YTHDF2表达、其临床病理特征和预后价值。进行功能性实验（包括增殖、集落形成和划痕愈合实验）以评估YTHDF2敲低的OSCC细胞系中的致癌表型。通过OSCC细胞系和来源于接受选择性YTHDF2抑制剂的OSCC患者的患者来源类器官（PDOs）在体外检测抗肿瘤效应。
结果：与非肿瘤组织和正常细胞相比，YTHDF2在OSCC组织和细胞系中显著上调，P < 0.001。YTHDF2的过表达与OSCC的发生、进展和不良预后相关。YTHDF2敲低显著损害了OSCC细胞的增殖、迁移和肿瘤球形成。在功能上，YTHDF2与m6A修饰的p53转录本发生物理相互作用并促进其降解。抑制YTHDF2显著促进了p53的稳定性和蛋白表达。YTHDF2抑制剂阻碍了癌细胞的生长、迁移、集落形成、肿瘤球和类器官发育。
结论：我们的结果确认YTHDF2作为一种致癌性m6A阅读器，通过增加p53 mRNA降解促进OSCC进展。我们的发现表明，YTHDF2可作为OSCC中一种候选抗肿瘤方法，在治疗上和肿瘤抑制方面加以利用。
查看英文原文 English abstract
Background: The N6-methyladenosine (m6A) RNA modification machinery has emerged as a critical modulator in cancer biology, however, the function of m6A “reader” proteins in oral squamous cell carcinoma (OSCC) is not fully understand. YTH domain family protein 2 (YTHDF2) is a prominent m6A-binding protein (m6A “reader”) that facilitates the degradation of methylated mRNAs and has been associated with tumorigenesis by targeting destabilization on clusters of tumor suppressor mRNAs. Nevertheless, the exact mechanism how YTHDF2 promotes OSCC proliferation and regulates p53 activity is in dire lack of elucidation. This study investigated the oncogenic role of YTHDF2 in OSCC and evaluated its potential as both a diagnostic biomarker and a therapeutic target.
Methods: The mRNA and protein levels of YTHDF2 were examined in OSCC tissues, matched adjacent non-cancerous specimens as well as OSCC cell lines by quantitative real-time PCR (qRT-PCR) and Western blot. Furthermore, the TCGA-OSCC dataset were employed to evaluate YTHDF2 expression, its clinicopathological characteristics and prognostic value. Functional assays including proliferation, colony formation and wound-healing assays were performed to evaluate the oncogenic phenotypes in YTHDF2-depleted OSCC cell lines. The antitumor effects were examined in vitro via OSCC cell lines and patient-derived organoids (PDOs) derived from OSCC patients who received a selective YTHDF2 inhibitor.
Results: YTHDF2 was remarkably up-regulated in OSCC tissues and cell lines compared with non-tumor tissues and normal cells, P < 0.001. Overexpression of YTHDF2 is correlated with OSCC development, progression and poor prognosis. YTHDF2 knockdown dramatically impaired OSCC cell proliferation, migration and tumorsphere formation. Functionally, YTHDF2 physically interacted with m6A-modified p53 transcripts and promoted their degradation. Inhibition of YTHDF2 significantly promoted the stability and protein expression of p53. YTHDF2 inhibitor impeded growth, migration, colony formation, tumorsphere and organoid development of cancer cells.
Conclusions: Our results recognize YTHDF2 as an oncogenic m6A reader promoting the OSCC progression by increasing p53 mRNA degradation. Our findings indicate that YTHDF2 can be exploited therapeutically and tumor suppressively as a candidate antitumor approach in OSCC.
利益披露 Disclosure
A. Pandi, None..
V. P. Jayaseelan, None..
P. Arumugam, None.