目的 生物信息学分析LncRNA NEAT1/WTAP/MYC调控轴，并通过敲降骨肉瘤细胞中LncRNA NEAT1，探究其对细胞增殖、迁移、侵袭及凋亡的影响，并检测该信号轴关键蛋白（WTAP和MYC）的表达变化。 方法 基于TARGET数据库筛选骨肉瘤枢纽基因，STRING构建蛋白互作网络，Starbase预测NEAT1-miRNAs结合位点及WTAP/MYC调控网络；合成si-NEAT1转染143B细胞，RT-qPCR验证敲降效率，MTT检测增殖、Transwell检测迁移/侵袭、流式检测凋亡、Western blot检测WTAP/MYC蛋白表达。 结果 生信分析揭示NEAT1作为ceRNA吸附miR-23a-3p/miR-150-5p上调WTAP，进而促进MYC表达；敲降NEAT1显著抑制骨肉瘤细胞增殖（P<0.001）、迁移/侵袭（P<0.001）并促进凋亡（P<0.001）；WB证实敲降后WTAP/MYC蛋白显著下调（P<0.01）。 结论 敲降LncRNA NEAT1可抑制骨肉瘤细胞增殖、迁移、侵袭并诱导细胞凋亡，其机制可能与LncRNA NEAT1调控WTAP和MYC的表达有关，本研究为骨肉瘤的机制研究和靶向治疗提供了理论依据。
Abstract
Objective To analyze the LncRNA NEAT1/WTAP/MYC regulatory axis through bioinformatics analysis, and by knockdown LncRNA NEAT1 in osteosarcoma cells, explore its effects on cell proliferation, migration, invasion and apoptosis, and simultaneously detect the expression changes of key proteins (WTAP and MYC) of this signal axis. Methods Osteosarcoma hub genes were screened based on the TARGET database, the protein-protein interaction network was constructed by STRING, and the binding sites of NEAT1-miRNAs and the WTAP/MYC regulatory network were predicted by Starbase. si-NEAT1 was synthesized and transfected into 143B cells. After the knockdown efficiency was verified by RT-qPCR，proliferation was detected by MTT, migration/invasion was detected by Transwell, apoptosis was detected by flow cytometry, and the expression of WTAP/MYC proteins was detected by Western blot. Results Bioinformatics analysis revealed that NEAT1, as ceRNA, adsorbed miR-23a-3p/miR-150-5p to up-regulate WTAP, thereby promoting the expression of MYC; Knockdown of NEAT1 significantly inhibited the proliferation of osteosarcoma cells (P<0.001), migration/invasion (P<0.001), and promoted apoptosis (P<0.001); WB confirmed WTAP/MYC protein was significantly downregulated after knockdown (P<0.01). Conclusions Knockdown of LncRNA NEAT1 can inhibit the proliferation, migration, invasion of osteosarcoma cells and induce cell apoptosis. The mechanism may be related to the regulation of WTAP and MYC expression by LncRNA NEAT1. This study provides a theoretical basis for the mechanism research and targeted therapy of osteosarcoma.
关键词
WTAP /
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LncRNA NEAT1 /
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骨肉瘤 /
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MYC /
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SiRNA
Key words
WTAP /
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LncRNA NEAT1 /
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Osteosarcoma /
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MYC /
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SiRNA
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参考文献
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基金
河南省科技攻关项目（LHGJ20200414）；郑州市科技创新指导计划项目（2025YLZDJH184）
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