首页|METTL3介导的PDK1 mRNA m6A修饰通过Akt/mTOR信号通路促进肺上皮细胞增殖

METTL3介导的PDK1 mRNA m6A修饰通过Akt/mTOR信号通路促进肺上皮细胞增殖

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腺苷N6-位点甲基化(m6A)在细胞增殖过程中发挥重要作用。RNA甲基转移酶3(METTL3)作为催化m6A关键酶，其介导m6A修饰在肺上皮细胞增殖中的作用机制尚不明确。本研究旨在探讨METTL3介导m6A修饰调控肺上皮细胞增殖的效应及机制。结果显示，在肺上皮细胞中敲低METTL3显著抑制细胞生长，而过表达METTL3则促进了细胞增殖(P＜0。05)。进一步的蛋白质免疫印迹结果显示，细胞生长和增殖的关键蛋白质PCNA在METTL3敲降的肺上皮细胞中蛋白质水平的表达显著下调，并且Akt以及mTOR的磷酸化水平显著降低(P＜0。05)。细胞免疫荧光结果发现，METTL3敲降的肺上皮细胞中m6A修饰水平显著降低(P＜0。05)。实时荧光定量PCR及蛋白质免疫印迹结果表明，Akt-mTOR信号通路上游调控分子PDK1的mRNA和蛋白质表达水平在METTL3敲降的肺上皮细胞中显著下降(P＜0。05)。机制上，m6A-IP-qPCR和RIP-qPCR结果进一步表明，METTL3催化PDK1 mRNA的3'UTR区域m6A修饰，进而被YTH N6-甲基腺苷RNA结合蛋白1(YTHDF1)识别，增强其mRNA的稳定性。总之，本研究揭示了 METTL3通过增强PDK1 m6A修饰，进而激活Akt-mTOR信号通路，促进细胞增殖。本研究为METTL3在上皮细胞增殖中的新角色提供了证据，同时为治疗肺上皮细胞损伤修复提供了新的治疗靶点。
METTL3-mediated m6A Modification of PDK1 mRNA Improves Lung Epithelial Cell Proliferation via Akt/mTOR Signaling
Adenosine N6-methylation(m6A)has been shown to be associated with the cell proliferation.The role of RNA methyltransferase 3(METTL3),a key enzyme catalyzing m6A,in mediating m6A modification in lung epi-thelial cell proliferation remains unclear.This study aims to explore the effects and mechanisms of METTL3-media-ted m6A modification in regulating lung epithelial cell proliferation.Results showed that knockdown of METTL3 sig-nificantly inhibited cell growth in lung epithelial cells,while overexpression of METTL3 promoted cell proliferation(P＜0.05).Further Western blotting results demonstrated that the expression levels of the key proteins PCNA in-volved in cell growth and proliferation were significantly downregulated in METTL3 knockdown lung epithelial cells,along with a significant decrease in phosphorylation levels of Akt and mTOR(P＜0.05).Immunofluorescence stai-ning revealed a significant decrease in m6A modification levels in METTL3 knockdown lung epithelial cells(P＜0.05).Real-time quantitative PCR and Western blotting results indicated that the mRNA and protein expression levels of PDK1,an upstream regulator of the Akt-mTOR signaling pathway,were significantly decreased in MET-TL3 knockdown lung epithelial cells(P＜0.05).Mechanistically,m6A-IP-qPCR and RIP-qPCR results further demonstrated that METTL3 catalyzed m6A modification in the 3'UTR region of PDK1 mRNA,which was then rec-ognized by YTH N6-methyladenosine RNA-binding protein 1(YTHDF1),enhancing the stability of its mRNA.In conclusion,this study reveals that METTL3 promotes cell proliferation by enhancing PDK1 m6 A modification,thereby activating the Akt-mTOR signaling pathway.It provides evidence for a novel role of METTL3 in epithelial cell proliferation and offers new therapeutic targets for lung epithelial cell injury repair.

methyltransferase 3(METTL3)RNA m6A modificationcell proliferationlung epithelial cellsphosphate inositol dependent protein kinase-1(PDK1)

靳艾、李梦宇、孙青竹

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山西医科大学公共卫生学院营养与食品卫生学教研室,太原 030001

山西医科大学营养与食品科学研究所,太原 030001

山西医科大学煤炭环境致病与防治教育部重点实验室,太原 030001

山西医科大学黄河流域生态公共卫生安全研究中心

西北农林科技大学动物科技学院动物科学系,陕西杨凌 712100

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甲基转移酶3 RNA m6A修饰 细胞增殖 肺上皮细胞 磷酸肌醇依赖性蛋白激酶-1

国家自然科学基金山西省基础研究计划山西省高等学校科技创新计划项目

823041462023030212121252023L061

2024

10.13865/j.cnki.cjbmb.2024.06.1035

中国生物化学与分子生物学报
中国生物化学与分子生物学会 北京大学

中国生物化学与分子生物学报

CSTPCD北大核心
影响因子：0.617
ISSN：1007-7626
年,卷(期)：2024.40(7)
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