Fourth Department of Orthopedic Surgery, Central Hospital Affiliated to Shenyang Medical College, No. 5 South Seven West Road, Tiexi, Shenyang, 110024, Liaoning, P.R. China.
Department of Cardiology, Shenyang Fourth People's Hospital, China Medical University, Shenyang, 110031, Liaoning, P.R. China.
Mol Biotechnol. 2024 Dec;66(12):3538-3548. doi: 10.1007/s12033-023-00953-2. Epub 2023 Oct 28.
As one of the most ubiquitous types of posttranscriptional modification, N6-methyladenosine (m6A) is extensively implicated in almost all types of cancers, including osteosarcoma. Our previous research partially uncovered the role of Metastasis Associated Lung Adenocarcinoma Transcript 1 (MALAT1) in osteosarcoma. However, the relationships between methyltransferase-like 3 (METTL3) and noncoding RNAs modified by METTL3, especially MALAT1, in osteosarcoma remain obscure.
The expression of METTL3 in osteosarcoma was evaluated by online bioinformatics analysis, immunohistochemical (IHC) staining, western blotting (WB), and reverse transcription-quantitative PCR (RT‒qPCR). Cell Counting Kit 8 (CCK-8) and Transwell assays were used to evaluate the cell proliferation and invasion abilities. The expression of MALAT1 in osteosarcoma was evaluated by online bioinformatics analysis and RT‒qPCR analysis. m6A methylated RNA immunoprecipitation-qPCR (MeRIP-qPCR) was used to detect m6A modification changes in MALAT1. An actinomycin D assay was used to study changes in the stability of MALAT1.
METTL3 was upregulated in osteosarcoma tissues and cell lines. Functionally, METTL3 promoted the proliferation and migration of osteosarcoma cells. Moreover, a clear positive correlation was found between METTL3 and MALAT1 expression, and MALAT1 was upregulated in osteosarcoma tissues and cells. Mechanistically, the presence of m6A modification sites in MALAT1 and METTL3-mediated m6A modification increased the stability of MALAT1 in osteosarcoma cells and promoted their proliferation and migration.
In this study, it was concluded that in osteosarcoma cells, METTL3, acting as an oncogene, promoted m6A modification of MALAT1, increased the stability of MALAT, and enhanced MALAT1-mediated oncogenic function.
作为最普遍的转录后修饰类型之一，N6-甲基腺苷（m6A）广泛参与几乎所有类型的癌症，包括骨肉瘤。我们之前的研究部分揭示了转移相关肺腺癌转录物 1（MALAT1）在骨肉瘤中的作用。然而，甲基转移酶样 3（METTL3）与 METTL3 修饰的非编码 RNA 之间的关系，特别是 MALAT1，在骨肉瘤中仍然不清楚。
通过在线生物信息学分析、免疫组织化学（IHC）染色、western blot（WB）和逆转录定量 PCR（RT-qPCR）评估骨肉瘤中 METTL3 的表达。细胞计数试剂盒 8（CCK-8）和 Transwell 测定用于评估细胞增殖和侵袭能力。通过在线生物信息学分析和 RT-qPCR 分析评估骨肉瘤中 MALAT1 的表达。m6A 修饰 RNA 免疫沉淀-qPCR（MeRIP-qPCR）用于检测 MALAT1 中的 m6A 修饰变化。放线菌素 D 测定用于研究 MALAT1 稳定性的变化。
METTL3 在骨肉瘤组织和细胞系中上调。功能上，METTL3 促进骨肉瘤细胞的增殖和迁移。此外，METTL3 和 MALAT1 表达之间存在明显的正相关，并且在骨肉瘤组织和细胞中 MALAT1 上调。机制上，MALAT1 中存在 m6A 修饰位点和 METTL3 介导的 m6A 修饰增加了骨肉瘤细胞中 MALAT1 的稳定性，并促进了其增殖和迁移。
在这项研究中，结论是在骨肉瘤细胞中，作为癌基因的 METTL3 促进 MALAT1 的 m6A 修饰，增加 MALAT1 的稳定性，并增强 MALAT1 介导的致癌功能。
