Zhao Hao, Li Jun, Yan Xin, Bian Xinchao
Department of Neurosurgery, Zibo Central Hospital, Zibo City, Shandong Province, 255036, People's Republic of China.
Department of Ultrasound, Central Hospital Affiliated to Shandong University, Zibo City, Shandong Province, 255036, People's Republic of China.
Cancer Manag Res. 2021 Apr 22;13:3493-3501. doi: 10.2147/CMAR.S274615. eCollection 2021.
LncRNA MAFG-AS1 plays critical roles in several types of cancer, while its role in glioblastoma (GBM) is unknown. By analyzing the TCGA dataset, we observed the upregulation of MAFG-AS1 in GBM. This study aimed to investigate the involvement of MAFG-AS1 in GBM cancer.
The expression levels of MAFG-AS1, mature miR-34a, and miR-34a precursor in GBM and paired non-tumor tissues of 56 GBM patients were determined by RT-qPCR. Correlations are analyzed using linear regression. Overexpression of MAFG-AS1 was achieved in GBM cells, followed by measurement of the expression levels of mature miR-34a, miR-34a precursor, DICER and Drosha by RT-qPCR. The roles of MAFG-AS1 and miR-34a in regulating GBM cell proliferation were evaluated by CCK-8 assay. Flow cytometry was performed to explore the role of MAFG-AS1 and miR-34a in regulating the apoptosis and cell cycle of GBM cells. Cell scratch experiment was performed to determine the role of MAFG-AS1 and miR-34a in regulating the migration of GBM cells. Subcutaneous tumor animal model was used for in vivo study. The expressions levels of BCL-2 and caspase-3 were detected by Western blot analysis.
MAFG-AS1 was upregulated in GBM, while mature miR-34a was downregulated in GBM. Interestingly, MAFG-AS1 was inversely correlated with mature miR-34a but not miR-34a precursor across GBM tissues. In GBM tissues, the overexpression of MAFG-AS1 did not affect the expression levels of miR-34a precursor but reduced the expression levels of mature miR-34a. MAFG-AS1 promoted the expression of DICER and Drosha in GBM cells. Moreover, the overexpression of MAFG-AS1 promoted the proliferation of GBM cells and reduced the inhibitory effects of miR-34a on cell proliferation but did not affect cell cycle, apoptosis and migration. The overexpression of MAFG-AS1 promoted the progression of GBM in vivo by promoting the proliferation of GBM cells while miR-34a reversed the effect of overexpression of MAFG-AS1.
MAFG-AS1 may suppress the maturation of miR-34a to promote GBM cell proliferation.
长链非编码RNA MAFG-AS1在多种癌症中发挥关键作用，但其在胶质母细胞瘤（GBM）中的作用尚不清楚。通过分析TCGA数据集，我们观察到GBM中MAFG-AS1上调。本研究旨在探讨MAFG-AS1在GBM中的作用。
采用RT-qPCR检测56例GBM患者的GBM组织及配对的非肿瘤组织中MAFG-AS1、成熟miR-34a和miR-34a前体的表达水平。使用线性回归分析相关性。在GBM细胞中实现MAFG-AS1过表达，然后通过RT-qPCR检测成熟miR-34a、miR-34a前体、DICER和Drosha的表达水平。通过CCK-8实验评估MAFG-AS1和miR-34a在调节GBM细胞增殖中的作用。采用流式细胞术探讨MAFG-AS1和miR-34a在调节GBM细胞凋亡和细胞周期中的作用。进行细胞划痕实验以确定MAFG-AS1和miR-34a在调节GBM细胞迁移中的作用。使用皮下肿瘤动物模型进行体内研究。通过蛋白质免疫印迹分析检测BCL-2和caspase-3的表达水平。
GBM中MAFG-AS1上调，而成熟miR-34a下调。有趣的是，在GBM组织中，MAFG-AS1与成熟miR-34a呈负相关，但与miR-34a前体无相关性。在GBM组织中，MAFG-AS1过表达不影响miR-34a前体的表达水平，但降低了成熟miR-34a的表达水平。MAFG-AS1促进GBM细胞中DICER和Drosha的表达。此外，MAFG-AS1过表达促进GBM细胞增殖，降低miR-34a对细胞增殖的抑制作用，但不影响细胞周期、凋亡和迁移。MAFG-AS1过表达通过促进GBM细胞增殖促进GBM在体内进展，而miR-34a可逆转MAFG-AS1过表达的作用。
MAFG-AS1可能通过抑制miR-34a成熟促进GBM细胞增殖。
