| 陈星宇,李维民,龙勇,向城卫,张施远,刘应刚.沉默RPARP-AS1对GBM U251细胞增殖、迁移和凋亡的影响.[J].中南医学科学杂志.,2024,(3):338-343. |
| 沉默RPARP-AS1对GBM U251细胞增殖、迁移和凋亡的影响 |
| Effects of silencing RPARP-AS1 on proliferation, migration and apoptosis of GBM U251 cells |
| 投稿时间：2023-06-30 修订日期：2024-03-12 |
| DOI：10.15972/j.cnki.43-1509/r.2024.03.006 |
| 中文关键词: RPARP-AS1 miR-339-5p MDM2 胶质母细胞瘤 细胞凋亡 细胞增殖 细胞迁移 [ |
| 英文关键词:RPARP-AS1 miR-339-5p MDM2 GBM cell apoptosis cell proliferation cell migration |
| 基金项目:四川省医学(青年创新)科研课题(Q21028) |
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| 中文摘要: |
| 目的探究沉默长链非编码RNA RPARP-AS1对胶质母细胞瘤(GBM)U251细胞增殖、迁移和凋亡的影响。 方法选取80例GBM组织与癌旁组织。将GBM细胞株U251细胞随机分为Control组、小干扰RNA阴性对照(si-NC)组、si-RPARP-AS1组、si-RPARP-AS1+抑制剂(inhibitor)NC组、si-RPARP-AS1+miR-339-5p inhibitor组。qRT-PCR检测组织和细胞RPARP-AS1、miR-339-5p和鼠双微体基因2(MDM2)mRNA的表达。CCK-8检测各组细胞增殖能力。Transwell检测各组U251细胞迁移和侵袭能力。流式细胞术检测细胞凋亡率。Western blotting检测B细胞淋巴瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)和MDM2蛋白的表达。双荧光素酶报告基因实验分别验证RPARP-AS1和miR-339-5p、miR-339-5p和MDM2的关系。 结果与癌旁组织相比,GBM组织RPARP-AS1、MDM2 mRNA高表达,miR-339-5p低表达(P＜0.05)。与Control组、si-NC组相比,si-RPARP-AS1组U251细胞RPARP-AS1、MDM2 mRNA和Bcl-2、N-cadherin、MDM2蛋白表达以及细胞增殖、迁移、侵袭能力显著降低(P＜0.05),miR-339-5p、Bax、E-cadherin蛋白表达以及细胞凋亡率显著升高(P＜0.05)。与si-RPARP-AS1组、si-RPARP-AS1+inhibitor NC组比较,si-RPARP-AS1+miR-339-5p inhibitor组细胞MDM2 mRNA和Bcl-2、N-cadherin、MDM2蛋白表达以及细胞增殖、迁移、侵袭能力显著升高(P＜0.05),miR-339-5p、Bax、E-cadherin蛋白表达以及细胞凋亡率显著降低(P＜0.05)。RPARP-AS1靶向负调控miR-339-5p表达,miR-339-5p靶向负调控MDM2表达。 结论沉默RPARP-AS1对GBM U251细胞增殖、迁移和侵袭具有抑制作用,对细胞凋亡具有促进作用,可能与通过上调miR-339-5p,抑制MDM2表达有关。 |
| 英文摘要: |
| AimTo investigate the effect of silencing long non-coding RNA RPARP-AS1 on the proliferation, migration and apoptosis of glioblastoma (GBM) U251 cells. Methods80 tumor and paracancerous tissue samples were collected from patients diagnosed with GBM and GBM cell line U251 was routinely cultured. U251 cells were randomly separated into Control group, small interfering RNA negative control (si-NC) group, si-RPARP-AS1 group, si-RPARP-AS1+inhibitor NC group, and si-RPARP-AS1+miR-339-5p inhibitor group. qRT-PCR method was applied to detect the expression of RPARP-AS1, miR-339-5p, and murine double minute 2 (MDM2) mRNA in tissues and cells. CCK-8 method was applied to detect the proliferation ability of cells in each group. Transwell method was applied to evaluate the migration and invasion abilities of U251 cell in each group. Flow cytometry was applied to detect cell apoptosis rate. Western blotting was applied to detect the expression of B cell lymphoblastoma-2 (Bcl-2), Bcl-2 associated X protein (Bax), E-cadherin, N-cadherin, and MDM2 proteins. Double luciferase reporter gene experiment was applied to verify the relationship between RPARP-AS1 and miR-339-5p, miR-339-5p and MDM2 respectively. ResultsCompared with adjacent cancer tissues, RPARP-AS1 and MDM2 mRNA were highly expressed in GBM tumor tissues, while miR-339-5p was lowly expressed (P＜0.05). Compared with the Control group and the si-NC group, the expression of RPARP-AS1, MDM2 mRNA, and Bcl-2, N-cadherin, MDM2 proteins, and proliferation value, migration, invasion ability in U251 cells of the si-RPARP-AS1 group were obviously decreased (P＜0.05). And the expression of miR-339-5p, Bax, E-cadherin protein, and cell apoptosis rate were obviously increased (P＜0.05). Compared with the si-RPARP-AS1 group and the si-RPARP-AS1+inhibitor NC group, the expression of MDM2 mRNA, and Bcl-2, N-cadherin, and MDM2 proteins, and proliferation value, migration, cell invasion ability in the si-RPARP-AS1+miR-339-5p inhibitor group were obviously increased (P＜0.05), whereas the expression of miR-339-5p, Bax and E-cadherin proteins, and cell apoptosis rate were obviously decreased (P＜0.05). RPARP-AS1 targeted and negatively regulated miR-339-5p expression, while miR-339-5p targeted and negatively regulated MDM2 expression. ConclusionSilencing RPARP-AS1 has inhibitory effects on the proliferation, migration, and invasion of GBM U251 cells, and promotes cell apoptosis, which may be related to the upregulation of miR-339-5p and inhibition of MDM2 expression. |
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