長いノンコーディングRNA VIM-AS1のノックダウンは神経膠腫細胞の増殖と遊走を抑制し、miR-105-5pを標的としたWEE1の調節を介して細胞のアポトーシスを増加させる

Knockdown of long non-coding RNA VIM-AS1 inhibits glioma cell proliferation and migration, and increases the cell apoptosis via modulation of WEE1 targeted by miR-105-5p.

• S-T Suo
• P Gong
• X-J Peng
• D Niu
• Y-T Guo

抄録

目的:

膠芽腫を含む神経膠腫は、世界的に原発性脳腫瘍の主要な種類である。グリオーマ形成にはLncRNAが関与している。本研究では、神経膠腫におけるVIM-AS1/miR-105-5p/WEE1シグナル伝達のメカニズムを明らかにすることを目的としている。

OBJECTIVE: Glioma including glioblastoma is the main type of primary brain tumors worldwide. LncRNAs have participated in glioma formation. This study aims to investigate the underlying mechanism for VIM-AS1/miR-105-5p/WEE1 signaling in glioma.

患者と方法:

PATIENTS AND METHODS: The clinical tumors and adjacent tissues were collected from 24 patients with glioma in the Shang Luo Central Hospital. Then, the clinical samples were subjected to hematoxylin-eosin staining (H&E). VIM-AS1, miR-105-5p, and WEE1 levels were measured using real-time PCR. The protein levels of WEE1, Cyclin A1, PCNA, N-cadherin, Vimentin, and Bcl-2, E-cadherin, and Bax were analyzed using Western blot. The overall survival of glioma patients was evaluated using the Kaplan-Meier analysis. The interaction between VIM-AS1 and miR-105-5p was determined using RIP assay and Dual-Luciferase reporter assay, and the binding between miR-105-5p and WEE1 was also detected by Dual-Luciferase reporter assay. Cell proliferation, colony formation, cell cycle, apoptosis, and migration were confirmed using CCK-8, colony formation assay, flow cytometry, and transwell assay, respectively.

結果:

VIM-AS1はがん組織で上昇しており、高レベルのVIM-AS1は全生存率の低下と正の相関があった。その結果、VIM-AS1はmiR-105-5pに結合してダウンレギュレートすることが明らかになった。さらに、VIM-AS1のノックダウンはin vivoでの腫瘍増殖を有意に抑制した。VIM-AS1のノックダウン／miR-105-5pの過剰発現は、Cyclin A1、PCNA、Vimentin、N-カドヘリン、Bcl-2の発現を阻害し、BaxおよびE-カドヘリンの発現を増加させることで、神経膠腫細胞の増殖、コロニー形成および遊走を抑制し、細胞のアポトーシスを促進した。興味深いことに、VIM-AS1 の過剰発現は、グリオーマ細胞における miR-105-5p の腫瘍抑制作用を逆転させた。また、WEE1の発現はVIM-AS1とmiR-105-5pによって相乗的に制御されていた。その結果、VIM-AS1はWEE1のアップレギュレーションやmiR-105-5pのダウンレギュレーションを介してグリオーマの進行を促進した。

RESULTS: VIM-AS1 was elevated in cancer tissues, and high level of VIM-AS1 was positively correlated with poor overall survival. Then, VIM-AS1 could bind to and downregulate miR-105-5p. Furthermore, the knockdown of VIM-AS1 significantly suppressed tumor growth in vivo. The knockdown of VIM-AS1/overexpression of miR-105-5p inhibited glioma cell growth, colony formation, and migration, and enhanced the cell apoptosis by inhibiting expression of Cyclin A1, PCNA, Vimentin, N-cadherin, and Bcl-2, and by increasing the expression of Bax and E-cadherin. Interestingly, the overexpression of VIM-AS1 reversed the tumor-suppressing role of miR-105-5p in glioma cells. Besides, the expression of WEE1 was synergistically regulated by VIM-AS1 and miR-105-5p. Consequently, VIM-AS1 promoted glioma progression via upregulating WEE1 or downregulating miR-105-5p.

結論:

VIM-AS1/miR-105-5p/WEE1シグナル伝達は神経膠腫治療の有望なターゲットとなる可能性がある。

CONCLUSIONS: VIM-AS1/miR-105-5p/WEE1 signaling may be a promising target for glioma treatment.