Jiang J, Wang S, Meng Q H, Yu R, Wei S C, Wang J, Qu C C, Wang C W
Department of Neurosurgery, the Second Hospital of Shandong University, Jinan 250033, China.
Department of Laboratory Medicine Center, the Second Hospital of Shandong University, Jinan 250033, China.
Zhonghua Yi Xue Za Zhi. 2020 Jun 2;100(21):1634-1639. doi: 10.3760/cma.j.cn112137-20200227-00482.
To screen the different microRNAs in the serum exosomes of patients with malignant glioma, to explore the effect of non-coding microRNA-376b-3p (miR-376b-3p) on the proliferation, invasion and tumor vasculogenic mimicry of glioma cells, and to verify its targeting effect on HOXD10. HiSeq/MiSeq high-throughput sequencing was used to screen the different microRNA expression profiles, target genes and action pathways in the serum exosomes of patients with malignant glioma. Samples were used to evaluate the expression of candidate microRNAs in serum exosomes of high-grade gliomas. The effects of miR-376b-3p on the proliferation, invasion and angiogenesis of glioma cells were detected by MTT assay, Transwell migration assay and Matrigel vasculogenic mimicry assay. The mRNA and protein expression of HOXD10 were detected to evaluate the regulatory effect of miR-376b-3p on it. There were 144 different expression microRNAs in the serum exosomes between malignant glioma and the normal control. Focal adhesion and tumor protein polysaccharides were involved in the regulation of glioma enriched by KEGG(Kyoto Encyclopedia of Genes and Genomes). MiR-376b-3p was down regulated in malignant glioma, and AUC of malignant glioma was 0.85 (0.01). MTT test showed that the proliferation ability of miR-376b-3p inhibitor group was higher than that of the control group, and that of miR-376b-3p mimic group was lower than that of the control group. Transwell migration test showed that the number of transmembrane cells in miR-376b-3p inhibitor group was higher than that in NC inhibitor group, and the number of transmembrane cells in miR-376b-3p mimic group was lower than that in NC mimic group. The number of tubes of vasculogenic mimicry in miR-376b-3p mimic group was lower than that in NC mimic group. MiR-376b-3p inhibitor decreased the expression level of HOXD10 mRNA and protein, and miR-376b-3p mimic increased the expression level of HOXD10 mRNA and protein. MiR-376b-3p is down-regulated in the serum exosomes of malignant glioma patients. The up-regulated miR-376b-3p can reduce the proliferation and invasion of glioma cells, inhibit the formation of vasculogenic mimicry, and increase the expression of HOXD10, which is expected to inhibit the formation of two forms of angiogenesis at the same time. MiR-376b-3p may be a new therapeutic target of anti-angiogenesis for malignant glioma.
筛选恶性胶质瘤患者血清外泌体中不同的微小RNA，探讨非编码微小RNA-376b-3p（miR-376b-3p）对胶质瘤细胞增殖、侵袭及肿瘤血管生成拟态的影响，并验证其对HOXD10的靶向作用。采用HiSeq/MiSeq高通量测序技术筛选恶性胶质瘤患者血清外泌体中不同的微小RNA表达谱、靶基因及作用通路。采集样本评估高级别胶质瘤患者血清外泌体中候选微小RNA的表达情况。通过MTT法、Transwell迁移实验和基质胶血管生成拟态实验检测miR-376b-3p对胶质瘤细胞增殖、侵袭及血管生成的影响。检测HOXD10的mRNA和蛋白表达，评估miR-376b-3p对其的调控作用。恶性胶质瘤与正常对照血清外泌体中有144种不同表达的微小RNA。KEGG（京都基因与基因组百科全书）富集分析显示粘着斑和肿瘤蛋白多糖参与胶质瘤的调控。恶性胶质瘤中miR-376b-3p表达下调，恶性胶质瘤的曲线下面积为0.85（0.01）。MTT实验显示，miR-376b-3p抑制剂组细胞增殖能力高于对照组，miR-376b-3p模拟物组低于对照组。Transwell迁移实验显示，miR-376b-3p抑制剂组穿膜细胞数高于阴性对照抑制剂组，miR-376b-3p模拟物组低于阴性对照模拟物组。miR-376b-3p模拟物组血管生成拟态的管腔数低于阴性对照模拟物组。miR-376b-3p抑制剂降低HOXD10 mRNA和蛋白表达水平，miR-376b-3p模拟物增加HOXD10 mRNA和蛋白表达水平。恶性胶质瘤患者血清外泌体中miR-376b-3p表达下调。上调miR-376b-3p可降低胶质瘤细胞的增殖和侵袭能力，抑制血管生成拟态的形成，并增加HOXD10的表达，有望同时抑制两种血管生成形式。miR-376b-3p可能是恶性胶质瘤抗血管生成治疗的新靶点。
