Bao Xing-Na, Wang Shang-Wei, Li Yongfeng
Department of Laboratory Medicine, Jining No. 1 People's Hospital, Jining, Shandong, China.
Department of Neurology, Sishui Country People's Hospital, Jining, Shandong, China.
Arch Med Sci. 2020 Feb 2;20(3):863-875. doi: 10.5114/aoms.2020.92817. eCollection 2024.
Accumulating evidence has proved that long non-coding RNAs (lncRNAs) are involved in progression of glioma. Nevertheless, the role of TUBA4B in glioma remains unclear.
The expression of the target gene was measured by quantitative RT-PCR. The prognostic role of TUBA4B was analyzed by Meier survival analysis. Cell proliferation, colony formation, apoptosis, cell cycle, migration and invasion were detected by MTS, soft agar colony forming assay, flow cytometry, and transwell assay. The target interaction of the target gene was validated by the luciferase reporter assay, biotin pull-down assay, and RNA immunoprecipitation.
We found that the expression of TUBA4B was lower in glioma tissues and cells. Moreover, patients with a low TUBA4B expression level exhibited poorer prognosis than those with high TUBA4B expression. Meanwhile, ROC analysis revealed that TUBA4B had diagnostic value to distinguish tumor patients from the healthy population. Overexpression of TUBA4B prohibited the malignancy of glioma, such as inhibition of proliferation, decrease of colony formation, arrest of the cell cycle, decline of migration and invasion, and promotion of cell apoptosis. In addition, we found that TUBA4B directly interacted with miR-183 and negatively regulated the expression of miR-183. We also observed that SMAD4 was a downriver target of miR-183 and TUBA4B subsequently exerted its tumor-suppressive effects by coordinating the expression of SMAD4 in glioma.
This study revealed for the first time that TUBA4B could be a tumor suppressor gene in glioma by adjustment of the TUBA4B/miR-183/SMAD4 axis, which may provide a useful prognostic biomarker and promising therapeutic target for glioma treatment.
越来越多的证据表明长链非编码RNA（lncRNAs）参与了胶质瘤的进展。然而，TUBA4B在胶质瘤中的作用仍不清楚。
通过定量逆转录聚合酶链反应（qRT-PCR）检测靶基因的表达。采用Meier生存分析评估TUBA4B的预后作用。通过MTS法、软琼脂集落形成试验、流式细胞术和Transwell试验检测细胞增殖、集落形成、凋亡、细胞周期、迁移和侵袭。通过荧光素酶报告基因试验、生物素下拉试验和RNA免疫沉淀验证靶基因的相互作用。
我们发现TUBA4B在胶质瘤组织和细胞中的表达较低。此外，TUBA4B表达水平低的患者预后比高表达患者差。同时，ROC分析显示TUBA4B对区分肿瘤患者和健康人群具有诊断价值。TUBA4B的过表达抑制了胶质瘤的恶性程度，如抑制增殖、减少集落形成、使细胞周期停滞、降低迁移和侵袭能力以及促进细胞凋亡。此外，我们发现TUBA4B直接与miR-183相互作用并负向调节miR-183的表达。我们还观察到SMAD4是miR-183的下游靶点，TUBA4B随后通过协调胶质瘤中SMAD4的表达发挥其肿瘤抑制作用。
本研究首次揭示TUBA4B可能通过调节TUBA4B/miR-183/SMAD4轴成为胶质瘤中的肿瘤抑制基因，这可能为胶质瘤治疗提供一个有用的预后生物标志物和有前景的治疗靶点。
