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Knockdown of long non-coding RNA LINC00941 suppressed cell proliferation, colony-formation, and migration of human glioblastoma cell lines
Folia Neuropathologica ( IF 2 ) Pub Date : 2023-05-22 , DOI: 10.5114/fn.2023.126894 Yuan Wang 1 , Shuwei Wang 1 , Haibo Wang 1 , Di Zhao 2 , Haoliang Zhang 1 , Huan Liu 1 , Jianzhong Cui 1 , Kaijie Wang 1
Folia Neuropathologica ( IF 2 ) Pub Date : 2023-05-22 , DOI: 10.5114/fn.2023.126894 Yuan Wang 1 , Shuwei Wang 1 , Haibo Wang 1 , Di Zhao 2 , Haoliang Zhang 1 , Huan Liu 1 , Jianzhong Cui 1 , Kaijie Wang 1
Affiliation
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.
中文翻译：
长链非编码 RNA LINC00941 的敲低抑制了人胶质母细胞瘤细胞系的细胞增殖、集落形成和迁移
简介：
胶质母细胞瘤 (GBM) 是成人中最常见和致命的原发性脑肿瘤类型，由于其高侵袭性，GBM 的治疗仍然具有挑战性。这项工作旨在阐明 LINC00941 在 GBM 中的作用。
材料和方法：
使用 siRNA 敲低两种 GBM 细胞系 U251 和 U87-MG 中的 LINC00941 表达。检查了 LINC00941 敲低的细胞增殖和集落形成能力。使用流式细胞术评估敲低的细胞凋亡，并使用蛋白质印迹检查 Bax、Bcl-2、cleaved caspase-3 的水平以及 ERK 和 Akt 的磷酸化。使用 Transwell 实验研究了敲低的迁移和侵袭。
结果：
与非肿瘤组织相比，GBM 中 LINC00941 的表达显着升高 ( p < 0.01)。对表达数据的统计分析进一步揭示了LINC00941与miR-526b-5p之间的负相关性（r = 0.7494，p < 0.001）。通过 RNA 干扰，LINC00941 在 U251 和 U87-MG 中被成功敲除。敲低显着抑制了细胞增殖和形成集落的能力。流式细胞术分析证明，两种细胞系的敲低均导致凋亡细胞的百分比升高，同时 Bcl-2 显着降低，Bax 和 cleaved caspase-3 显着增加。通过敲低，两种细胞系中 ERK 和 Akt 的磷酸化也得到增强。此外，LINC00941 的敲低抑制了两种细胞系穿过 Transwell 膜和基质胶的迁移。
结论：
LINC00941在GBM中过表达，在细胞增殖和存活、迁移和侵袭中发挥重要作用。
更新日期：2023-05-22
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.
中文翻译：
长链非编码 RNA LINC00941 的敲低抑制了人胶质母细胞瘤细胞系的细胞增殖、集落形成和迁移
简介：
胶质母细胞瘤 (GBM) 是成人中最常见和致命的原发性脑肿瘤类型，由于其高侵袭性，GBM 的治疗仍然具有挑战性。这项工作旨在阐明 LINC00941 在 GBM 中的作用。
材料和方法：
使用 siRNA 敲低两种 GBM 细胞系 U251 和 U87-MG 中的 LINC00941 表达。检查了 LINC00941 敲低的细胞增殖和集落形成能力。使用流式细胞术评估敲低的细胞凋亡，并使用蛋白质印迹检查 Bax、Bcl-2、cleaved caspase-3 的水平以及 ERK 和 Akt 的磷酸化。使用 Transwell 实验研究了敲低的迁移和侵袭。
结果：
与非肿瘤组织相比，GBM 中 LINC00941 的表达显着升高 ( p < 0.01)。对表达数据的统计分析进一步揭示了LINC00941与miR-526b-5p之间的负相关性（r = 0.7494，p < 0.001）。通过 RNA 干扰，LINC00941 在 U251 和 U87-MG 中被成功敲除。敲低显着抑制了细胞增殖和形成集落的能力。流式细胞术分析证明，两种细胞系的敲低均导致凋亡细胞的百分比升高，同时 Bcl-2 显着降低，Bax 和 cleaved caspase-3 显着增加。通过敲低，两种细胞系中 ERK 和 Akt 的磷酸化也得到增强。此外，LINC00941 的敲低抑制了两种细胞系穿过 Transwell 膜和基质胶的迁移。
结论：
LINC00941在GBM中过表达，在细胞增殖和存活、迁移和侵袭中发挥重要作用。
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