Department of Neurosurgery, The First Hospital of Handan, Handan, Hebei Province, China.
Department of E.N.T, The First Hospital of Handan, Handan, Hebei Province, China.
Aging (Albany NY). 2021 Nov 11;13(21):24339-24348. doi: 10.18632/aging.203684.
In the present study, we found the expression of miR-15a-5p (miR-15a) was increased in glioma tissues, and we further explore the underlying mechanism of miR-15a in glioma progression. Microarray analysis used to identify the differentially expressed microRNAs (miRNAs) in glioma tissues. The expression of miR-15a in glioma tissues and cell lines was tested by qRT-PCR. Luciferase assay was used to determine the binding between miR-15a and Smad7. Wound healing and transwell assay were used to examine the role of miR-15a/Smad7 in SHG139 cells. Western blot was used to detect the protein level of Smad7 and epithelial-mesenchymal transition (EMT) markers. A tumor formation model in nude mice was established to measure the role of miR-15a . MiR-15a was significantly increased in glioma tissues and cells, which indicated a poor prognosis of glioma patients. MiR-15a mimics induced miR-15a level in SHG139 cells, and promoted the malignancy of SHG139 cells, while miR-15a inhibitor showed the opposite effects. Luciferase assay indicated that Smad7 was the direct target of miR-15a, and Smad7 was down-regulated in glioma tissues. Functional experiments revealed that miR-15a inhibitor inhibited the EMT pathway and the migration and invasion of glioma cells, but the silencing of Smad7 reversed the effects of miR-15a inhibitor in EMT pathway and glioma progression. Finally, we performed animal experiments to verify the role of miR-15a . Present study showed that deletion of miR-15a inhibited the activation of EMT signaling via targeting Smad7, thus suppressed the tumorigenesis and tumor growth of glioma.
在本研究中，我们发现 miR-15a-5p（miR-15a）在胶质瘤组织中的表达增加，我们进一步探讨了 miR-15a 在胶质瘤进展中的潜在机制。使用微阵列分析鉴定胶质瘤组织中差异表达的 microRNAs（miRNAs）。通过 qRT-PCR 检测 miR-15a 在胶质瘤组织和细胞系中的表达。使用荧光素酶报告基因实验确定 miR-15a 与 Smad7 之间的结合。通过划痕愈合和 Transwell 实验检测 miR-15a/Smad7 在 SHG139 细胞中的作用。Western blot 用于检测 Smad7 和上皮间质转化（EMT）标志物的蛋白水平。建立裸鼠肿瘤形成模型以测量 miR-15a 的作用。miR-15a 在胶质瘤组织和细胞中显著增加，这表明胶质瘤患者的预后不良。miR-15a 模拟物诱导 SHG139 细胞中 miR-15a 水平升高，并促进 SHG139 细胞的恶性转化，而 miR-15a 抑制剂则显示出相反的效果。荧光素酶报告基因实验表明 Smad7 是 miR-15a 的直接靶标，并且 Smad7 在胶质瘤组织中下调。功能实验表明 miR-15a 抑制剂抑制 EMT 通路以及胶质瘤细胞的迁移和侵袭，但是 Smad7 的沉默逆转了 miR-15a 抑制剂在 EMT 通路和胶质瘤进展中的作用。最后，我们进行了动物实验以验证 miR-15a 的作用。本研究表明，缺失 miR-15a 通过靶向 Smad7 抑制 EMT 信号的激活，从而抑制胶质瘤的肿瘤发生和肿瘤生长。
