METTL3 介导的 pri-miR-148a-3p 的 m6A 修饰通过调节 TXNIP 影响前列腺癌的进展。
METTL3-mediated m6A modification of pri-miR-148a-3p affects prostate cancer progression by regulating TXNIP.
机构信息
Department of Urology, The Second Xiangya Hospital, Central South University, Changsha, China.
Department of Histology and Embryology, Xiangya School of Medicine, Central South University, Changsha, China.
出版信息
Environ Toxicol. 2023 Oct;38(10):2377-2390. doi: 10.1002/tox.23874. Epub 2023 Jul 14.
OBJECTIVE
Prostate cancer (PCa) severely affects men's health worldwide. The mechanism of methyltransferase-like 3 (METTL3) in affecting PCa development by regulating miR-148a-3p expression via N6-methyladenosine (m6A) modification was investigated.
METHODS
METTL3, miR-148a-3p, and thioredoxin interacting protein (TXNIP) levels were determined using RT-qPCR and Western blotting. The m6A modification level of miR-148a-3p was observed by Me-RIP assay. Bioinformatics website predicted miR-148a-3p and TXNIP levels in PCa and their correlation, and the binding site between them was verified by dual-luciferase assay. The proliferation, migration, invasion, and apoptosis of PCa cells were examined by CCK-8 assay, Transwell assay, and flow cytometry. A transplanted tumor model was established in nude mice to observe the tumor growth ability, followed by determination of TXNIP levels in tumor tissues by immunohistochemistry.
RESULTS
METTL3 interference restrained the proliferation, migration, and invasion and promoted apoptosis of PCa cells. METTL3 up-regulated miR-148a-3p by promoting the m6A modification of pri-miR-148a-3p in PCa cells. miR-148a-3p overexpression nullified the inhibitory actions of silencing METTL3 on PCa cell growth. miR-148a-3p facilitated PCa cell growth by silencing TXNIP. METTL3 interference inhibited tumor growth by down-regulating miR-148a-3p and up-regulating TXNIP.
CONCLUSION
METTL3 promoted miR-148a-3p by mediating the m6A modification of pri-miR-148a-3p, thereby targeting TXNIP, interfering with METTL3 to inhibit the proliferation, migration and invasion of PCa cells, promote apoptosis, and inhibit tumor growth in nude mice.
目的
前列腺癌（PCa）严重影响全球男性健康。本研究旨在探讨甲基转移酶样 3（METTL3）通过调节miR-148a-3p 的表达来影响 PCa 发展的机制，即通过 N6-甲基腺苷（m6A）修饰来影响 miR-148a-3p 的表达。
方法
采用 RT-qPCR 和 Western blot 检测 METTL3、miR-148a-3p 和硫氧还蛋白相互作用蛋白（TXNIP）的水平。采用 Me-RIP assay 观察 miR-148a-3p 的 m6A 修饰水平。生物信息学网站预测 PCa 中 miR-148a-3p 和 TXNIP 的水平及其相关性，并通过双荧光素酶报告基因实验验证它们之间的结合位点。采用 CCK-8 assay、Transwell assay 和流式细胞术检测 PCa 细胞的增殖、迁移和侵袭以及凋亡。建立裸鼠移植瘤模型观察肿瘤生长能力，然后采用免疫组化法检测肿瘤组织中 TXNIP 的水平。
结果
干扰 METTL3 抑制了 PCa 细胞的增殖、迁移和侵袭，促进了其凋亡。METTL3 通过促进 pri-miR-148a-3p 的 m6A 修饰，在 PCa 细胞中上调了 miR-148a-3p。沉默 METTL3 对 PCa 细胞生长的抑制作用被 miR-148a-3p 的过表达所消除。miR-148a-3p 通过沉默 TXNIP 促进了 PCa 细胞的生长。干扰 METTL3 通过下调 miR-148a-3p 和上调 TXNIP 抑制了肿瘤的生长。
结论
METTL3 通过介导 pri-miR-148a-3p 的 m6A 修饰促进了 miR-148a-3p 的表达，从而靶向 TXNIP，干扰 METTL3 抑制了 PCa 细胞的增殖、迁移和侵袭，促进了其凋亡，并抑制了裸鼠肿瘤的生长。
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