| 摘要: |
| ［摘要］ 目的 探讨环状RNA(CircRNA)SEPT9调节miR-650/热休克蛋白B1(HSPB1)轴对胶质瘤细胞恶性生物学行为的影响。方法 通过实时定量聚合酶链式反应（qRT-PCR）、Western blot法检测胶质瘤组织、癌旁组织、人类正常神经胶质细胞HEB及胶质瘤细胞系LN18、SW1088、U251中CircSEPT9、miR-650、HSPB1蛋白表达水平。将U251细胞分为对照组（NC组）、si-NC组、si-CircSEPT9组、mimic NC组、miR-650 mimic组、si-CircSEPT9+inhibitor NC组、si-CircSEPT9+miR-650 inhibitor组、si-CircSEPT9+pcDNA组、si-CircSEPT9+pcDNA-HSPB1组。采用qRT-PCR法检测CircSEPT9、miR-650表达水平。采用Western blot法检测HSPB1、细胞周期蛋白D1(Cyclin D1)、Caspase-3、基质金属蛋白酶（MMP）-2、MMP-9蛋白表达水平。通过双荧光素酶基因实验验证CircSEPT9与miR-650、miR-650与HSPB1的关系。结果 在胶质瘤组织和细胞中，CircSEPT9和HSPB1蛋白呈高表达，miR-650呈低表达，且在U251细胞中CircSEPT9以及HSPB1蛋白表达水平最高，miR-650表达水平最低，差异有统计学意义（P<0.05）。沉默CircSEPT9或过表达miR-650均可抑制U251细胞增殖、侵袭、迁移及Cyclin D1、MMP-2、MMP-9蛋白表达水平，促进细胞凋亡及Caspase-3蛋白表达。下调miR-650或过表达HSPB1均可减弱沉默CircSEPT9对U251细胞的作用影响。CircSEPT9与miR-650、miR-650与HSPB1存在靶向关系。结论 沉默CircSEPT9可能通过海绵化上调miR-650来抑制HSPB1表达，进而抑制U251细胞增殖、迁移与侵袭，促进细胞凋亡。 |
| 关键词: 环状RNA SEPT9 miR-650 热休克蛋白B1 胶质瘤 生物学行为 |
| DOI：10.3969/j.issn.1674-3806.2023.08.16 |
| 分类号:R 739.41 |
| 基金项目:河北省卫生健康委2021年度医学科学研究课题计划项目（编号：20211540） |
|
| An investigation of the effect of CircSEPT9 on malignant biological behavior of glioma cells by regulating miR-650/HSPB1 axis |
|
REN Cheng-bo, ZHAO Li-xia, DU Xiao-meng, et al.
|
|
Department of Radiotherapy, the First Affiliated Hospital of Hebei North University, Zhangjiakou 075000, China
|
| Abstract: |
| ［Abstract］ Objective To investigate the effect of circular ribonucleic acid(CircRNA) septin 9(CircSEPT9) on malignant biological behavior of glioma cells by regulating micro ribonucleic acid-650(miR-650)/heat shock protein B1(HSPB1) axis. Methods The expressions of CircSEPT9, miR-650 and HSPB1 protein in glioma tissues, tumor-adjacent tissues, human normal glial cells HEB and glioma cell lines LN18, SW1088 and U251 were detected by quantitative real time polymerase chain reaction(qRT-PCR) and Western blot, respectively. U251 cells were divided into control group(NC group), si-NC group, si-CircSEPT9 group, mimic NC group, miR-650 mimic group, si-CircSEPT9+inhibitor NC group, si-CircSEPT9+miR-650 inhibitor group, si-CircSEPT9+pcDNA group, and si-CircSEPT9+pcDNA-HSPB1 group. The expression levels of CircSEPT9 and miR-650 were detected by qRT-PCR. Western blot was applied to detect the expression levels of HSPB1, Cyclin D1, Caspase-3, matrix metalloproteinase(MMP)-2 and MMP-9 proteins. Double luciferase gene experiment was applied to verify the relationship between CircSEPT9 and miR-650, and the relationship between miR-650 and HSPB1. Results In the glioma tissues and glioma cells, CircSEPT9 and HSPB1 were highly expressed, while miR-650 was low expressed. In the U251 cells, the expression levels of CircSEPT9 and HSPB1 were the highest, and the expression level of miR-650 was the lowest, and the differences were significant(P<0.05). Silencing CircSEPT9 or overexpressing miR-650 was able to inhibit U251 cell proliferation, invasion, migration and the expression levels of Cyclin D1, MMP-2 and MMP-9 proteins, and was able to promote cell apoptosis and Caspase-3 protein expression. The down-regulation of miR-650 or overexpression of HSPB1 could weaken the effect of CircSEPT9 silencing on the U251 cells. CircSEPT9 had a targeting relationship with miR-650, and miR-650 had a targeting relationship with HSPB1. Conclusion Silencing CircSEPT9 may inhibit the expression of HSPB1 by up-regulating miR-650 through spongification, thereby inhibiting the proliferation, migration and invasion of U251 cells and promoting apoptosis. |
| Key words: Circular ribonucleic acid(CircRNA) septin 9(CircSEPT9) Micro ribonucleic acid-650(miR-650) Heat shock protein B1(HSPB1) Glioma Biological behavior |
