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长链非编码 RNA LINC00941 的敲低抑制了人胶质母细胞瘤细胞系的增殖、集落形成和迁移。

Knockdown of long non-coding RNA LINC00941 suppressed cell proliferation, colony-formation, and migration of human glioblastoma cell lines.

机构信息

Department of Neurosurgery, Tangshan Gongren Hospital, Tangshan, Hebei, China.

Department of Neurosurgery, Hebei Provincial Hospital of Traditional Chinese Medicine, Shijiazhuang, Hebei, China.

出版信息

Folia Neuropathol. 2023;61(2):209-216. doi: 10.5114/fn.2023.126894.

Abstract

INTRODUCTION

Glioblastoma (GBM) represents the most common and lethal type of primary brain tumour in adults, and due to its high invasiveness, treatment of GBM remains challenging. This work is aimed to elucidate the role of LINC00941 in GBM.

MATERIAL AND METHODS

Expression of LINC00941 in two GBM cell lines U251 and U87-MG was knocked down using siRNA. Cell proliferation and colony-formation ability of LINC00941 knockdown were examined. Apoptosis of the knockdown was evaluated using flow cytometry, with the levels of Bax, Bcl-2, cleaved caspase-3, and phosphorylation of ERK and Akt to be examined using western blotting. Migration and invasion of the knockdown was studied using transwell assays.

RESULTS

Expression of LINC00941 was significantly elevated in GBM compared to non-tumour tissues ( p < 0.01). Statistical analysis on the expression data further revealed the negative correlation between LINC00941 and miR-526b-5p ( r = 0.7494, p < 0.001). LINC00941 was successfully knocked down with RNA interference in U251 and U87-MG. The knockdown significantly suppressed cell proliferation and the ability to form colonies. Percentage of apoptotic cells was elevated by the knockdown in both cell lines as evidenced by flow cytometric analysis, which was accompanied by a significant decrease in Bcl-2 and substantial increases in Bax and cleaved caspase-3. Phosphorylation of ERK and Akt was also enhanced in both cell lines by the knockdown. In addition, knockdown of LINC00941 suppressed migration of both cell lines across transwell membrane and matrigel.

CONCLUSIONS

LINC00941 is overexpressed in GBM, exhibiting important roles in cell proliferation and survival, migration and invasion.

摘要

简介

胶质母细胞瘤(GBM)是成人中最常见和最致命的原发性脑肿瘤类型,由于其高度侵袭性,GBM 的治疗仍然具有挑战性。本工作旨在阐明 LINC00941 在 GBM 中的作用。

材料与方法

使用 siRNA 敲低两种 GBM 细胞系 U251 和 U87-MG 中的 LINC00941 表达。检查 LINC00941 敲低对细胞增殖和集落形成能力的影响。使用流式细胞术评估敲低对细胞凋亡的影响,使用 Western blot 检测 Bax、Bcl-2、裂解 caspase-3 的水平以及 ERK 和 Akt 的磷酸化。使用 Transwell 测定研究敲低对细胞迁移和侵袭的影响。

结果

与非肿瘤组织相比,GBM 中 LINC00941 的表达明显升高(p<0.01)。对表达数据的统计分析进一步显示 LINC00941 与 miR-526b-5p 之间呈负相关(r=0.7494,p<0.001)。在 U251 和 U87-MG 中成功使用 RNA 干扰敲低 LINC00941。敲低显著抑制细胞增殖和集落形成能力。流式细胞术分析表明,敲低导致两种细胞系中的凋亡细胞百分比升高,同时 Bcl-2 显著减少,Bax 和裂解 caspase-3 显著增加。敲低还增强了两种细胞系中 ERK 和 Akt 的磷酸化。此外,LINC00941 的敲低抑制了两种细胞系穿过 Transwell 膜和 Matrigel 的迁移。

结论

LINC00941 在 GBM 中过度表达,在细胞增殖和存活、迁移和侵袭中发挥重要作用。

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