Cell Biology and Toxicology ( IF 5.3 ) Pub Date : 2021-01-05 , DOI: 10.1007/s10565-020-09570-0 Peng Zhao 1 , Jianhua Cheng 1 , Bin Li 1 , Ding Nie 1 , Chuzhong Li 2 , Songbai Gui 1 , Hongyun Wang 2 , Yazhuo Zhang 2
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This study explored the function of microRNAs (miRNAs) in invasive pituitary adenomas (IPA), and developed a microRNA-exosome strategy for the disease treatment. Differentially expressed miRNAs and tumor-associated markers in IPA, non-invasive pituitary adenoma (NIPA), and rat pituitary adenoma cells were identified by bioinformatics analysis and qRT-PCR. Then, the cells were treated by miR-149-5p and miR-99a-3p mimics or inhibitors, or incubated with modified exosome with overexpressed or silenced miRNAs. The cell behaviors were analyzed by molecular experiments. Xenograft assays were constructed by injection of pituitary adenoma cells and exosome into NU/NU nude mice. Tumor size, weight, and expressions of markers related to miRNAs and angiogenesis were determined. Target genes for miR-99a-3p and miR-149 were predicted and verified by bioinformatics analysis and molecular experiments. Twenty differentially expressed miRNAs were identified, among which miR-99a-3p and miR-149 were inhibited in both pituitary adenoma cells and tissues significantly. Expressions of E-cadherin and p53 were down-regulated, while those of MMP-2, MMP-9, N-cadherin, Vimentin, and VEGF were up-regulated in pituitary adenoma cells and tissues, especially in IPA. Further experiments revealed that overexpressed miR-149 and miR-99a-3p inhibited the growth and metastasis of pituitary adenoma cells and tube formation of endothelial cells. MiR-149 and miR-99a-3p overexpressed by exosome showed similar suppressive effects on cell viability, metastasis, tube formation ability, in vivo tumor growth, and expressions of angiogenesis-related markers. Further analysis showed that NOVA1, DTL, and RAB27B were targeted by miR-99a-3p. This study found that overexpressed miR-149-5p and miR-99a-3p induced by exosome could suppress the progression of IPA.
Graphical abstract and graphical headlights
中文翻译：
上调外泌体 MiR-149-5p 和 MiR-99a-3p 的表达抑制垂体腺瘤的进展
本研究探讨了 microRNA (miRNA) 在侵袭性垂体腺瘤 (IPA) 中的功能，并开发了一种用于疾病治疗的 microRNA-外泌体策略。通过生物信息学分析和 qRT-PCR 鉴定 IPA、非侵袭性垂体腺瘤 (NIPA) 和大鼠垂体腺瘤细胞中差异表达的 miRNA 和肿瘤相关标志物。然后，用 miR-149-5p 和 miR-99a-3p 模拟物或抑制剂处理细胞，或与具有过表达或沉默 miRNA 的修饰外泌体一起孵育。通过分子实验分析细胞行为。通过将垂体腺瘤细胞和外泌体注射到 NU/NU 裸鼠中来构建异种移植试验。确定肿瘤大小、重量和与 miRNA 和血管生成相关的标志物的表达。通过生物信息学分析和分子实验预测和验证了 miR-99a-3p 和 miR-149 的靶基因。鉴定出20个差异表达的miRNA，其中miR-99a-3p和miR-149在垂体腺瘤细胞和组织中均被显着抑制。E-cadherin和p53的表达下调，而MMP-2、MMP-9、N-cadherin、Vimentin和VEGF在垂体腺瘤细胞和组织中的表达上调，尤其是在IPA中。进一步的实验表明，过表达的 miR-149 和 miR-99a-3p 抑制了垂体腺瘤细胞的生长和转移以及内皮细胞的管形成。外泌体过表达的 MiR-149 和 miR-99a-3p 对细胞活力、转移、管形成能力、体内肿瘤生长、. 进一步分析表明，NOVA1、DTL 和 RAB27B 被 miR-99a-3p 靶向。本研究发现外泌体诱导的过表达 miR-149-5p 和 miR-99a-3p 可以抑制 IPA 的进展。
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