| 吕旭阳,孙江川,董玥,等.miR-3175/SIX5轴调节神经胶质瘤细胞的增殖和迁移[J].浙江中医药大学学报,2022,46(1):40-49. |
| miR-3175/SIX5轴调节神经胶质瘤细胞的增殖和迁移 |
| MiR-3175/SIX5 Axis Regulated Cell Proliferation and Migration of Glioma Cells |
| DOI：10.16466/j.issn1005-5509.2022.01.007 |
| 中文关键词: miR-3175 胶质瘤 细胞增殖 细胞凋亡 细胞周期 sineoculis同源异型盒同源物5 增殖 迁移 |
| 英文关键词: miR-3175 glioma cell proliferation cell apoptosis cell cycle sineoculis homeobox homologue 5 proliferation migration |
| 基金项目:浙江省自然基金探索项目(LY20H280005)；国家自然科学基金青年项目(81602624) |
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| 中文摘要: |
| [目的] 探讨miR-3175在胶质瘤细胞中的表达，以及对胶质瘤细胞增殖和迁移能力的影响及相关作用机制。[方法] 采用实时荧光定量聚合酶链式反应(Real-time quantitative polymerase chain reaction，Real-time qPCR)检测胶质瘤细胞与人正常胶质细胞中miR-3175的表达水平。向胶质瘤细胞株中转染miR-3175 mimics和inhibitors后，采用噻唑蓝(3-(4，5-Dimethylthiazol-2-yl)-2， |
| 英文摘要: |
| [Objective] To investigate the expression of miR-3175 in glioma cells and its effect on the proliferation and migration of glioma cells and relevant mechanism. [Methods] The Real-time quantitative polymerase chain reaction(Real-time qPCR) was used to detect the expression of miR-3175 in glioma cells and human normal glial cells. After the miR-3175 mimics and inhibitors were transfected into A172 and U251 cells， 3-(4，5-Dimethylthiazol-2-yl)-2，5-diphenyltetrazolium bromide(MTT) assay was used to detect the proliferation of glioma cells in each group， the Transwell assay was used to detect the migration of cells， the flow cytometry was used to detect the apoptotic rate of cells. The target of miR-3175 was predicted by miRTarBase and TargetScan. Dual luciferase reporter gene system， Western blot and Real-time qPCR were used to detect the interaction between miR-3175 and sineoculis homeobox homologue 5(SIX5); bioinformatics and Western blot were used to analyze the expression of SIX5 in glioma. The pCDNA3.4-SIX5， pCDNA3.4-SIX5+miR-3175 mimics and miR-3175 mimics were transfected into glioma cells， MTT， Transwell and flow cytometry assays were performed to evaluate the effect of miR-3175 and its target gene SIX5 on the malignant biological behavior of glioma cells. [Results] The results of Real-time qPCR show that the expression level of miR-3175 is significantly lower in glioma cells than human normal glial cells(P＜0.01). The results of vitro transfection experiments show that the up-regulation of miR-3175 could significantly inhibit the proliferation and migration of glioma cells(P＜0.05)， and induce apoptosis by arresting the cell cycle of glioma cells in the G0/G1 phase(P＜0.05， P＜0.01). The prediction results of bioinformatics show that SIX5 is probably a potential target gene of miR-3175. The Western blot analysis and luciferase reporter gene assay results further verify that the expression of SIX5 is negatively correlated with miR-3175(P＜0.01). The results of vitro transfection experiments show that the proliferation and migration of glioma cells are significantly increased(P＜0.05， P＜0.01)， while the apoptotic rate of glioma cells is significantly reduced after transfected pCDNA3.4-SIX5(P＜0.05， P＜0.01)， overexpression of SIX5 can abrogate the inhibitory effect of miR-3175 on malignant biological behavior of glioma. [Conclusion] The expression level of miR-3175 is significantly downregulated in glioma， and miR-3175 inhibits the malignant biological behavior of glioma. The possible mechanism is to inhibit the expression of cancer promoting gene SIX5， so as to reduce the proliferation and migration ability of glioma， suggesting that it might have the potential to become a new target for glioma diagnosis and treatment. |
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