Jiang Shu, Luo Chao, Chen Yongli, Chen Jing, Tao Shuang, Zou Quan, He Chunzhi, Dong Shanwu
Department of Pediatrics, Wuhan Fourth Hospital, No. 76 Jiefang Avenue, Qiaokou District, Wuhan, 430034, China.
Biochem Genet. 2023 Apr;61(2):538-550. doi: 10.1007/s10528-022-10264-x. Epub 2022 Aug 19.
Glioblastoma (GBM) is the most malignant and challenging type of astrocytoma and also notoriously acknowledged as the most common primary brain tumor globally. Currently, chemotherapy is the most master therapy for tumor and is essential in clinical treatment for GBM. Nevertheless, the characterization of chemotherapy resistance seriously hinders clinical chemotherapy treatment. Accordingly, there are imperious demands for the exploitation of novel chemosensitizer to promote the efficacy of chemotherapy. Our current study was conducted to probe into the potential impacts of microRNA (miR)-640 on the chemosensitivity in GBM and the associated underlying mechanism. Initially, TargetScan software was utilized to predict the targeted genes of miR-640, and the target relationship between miR-640 and Bcl-2-modifying factor (BMF) was validated by double luciferase report assay. Additionally, to explore the role of miR-640/BMF in U251 cells, miR-640 inhibitor/BMF-siRNA was used. U251 cells were processed with 100 μM temozolomide (TMZ) and detected with CCK-8 kit. Eventually, RT-qPCR and Western blotting were used for evaluating Bcl-2, Bax mRNA, and protein expression level. Flow cytometry analysis was performed to measure cellular apoptosis. Initially, the results indicated that BMF was the target gene of miR-640. MiR-640 negatively regulated BMF expression in GBM cells. Besides, the findings revealed that miR-640 inhibition significantly inhibited U251 cell proliferation, promoted cell apoptosis, and increased the sensitivity of GBM cells to TMZ by targeting BMF. Moreover, BMF overexpression significantly suppressed U251 cell proliferation, induced cell apoptosis, and increased the sensitivity of GBM cells to TMZ. Inhibition of miR-640 expression enhances chemosensitivity of human GBM cells to TMZ by targeting BMF.
胶质母细胞瘤（GBM）是星形细胞瘤中最恶性且最具挑战性的类型，也是全球公认的最常见的原发性脑肿瘤。目前，化疗是治疗肿瘤的主要方法，在GBM的临床治疗中至关重要。然而，化疗耐药性的特征严重阻碍了临床化疗治疗。因此，迫切需要开发新型化学增敏剂以提高化疗效果。我们目前的研究旨在探讨微小RNA（miR）-640对GBM化疗敏感性的潜在影响及其相关潜在机制。首先，利用TargetScan软件预测miR-640的靶基因，并通过双荧光素酶报告基因检测验证miR-640与Bcl-2修饰因子（BMF）之间的靶向关系。此外，为了探究miR-640/BMF在U251细胞中的作用，使用了miR-640抑制剂/BMF小干扰RNA。用100μM替莫唑胺（TMZ）处理U251细胞，并用CCK-8试剂盒进行检测。最后，采用逆转录-定量聚合酶链反应（RT-qPCR）和蛋白质免疫印迹法评估Bcl-2、Bax的mRNA和蛋白表达水平。通过流式细胞术分析检测细胞凋亡情况。首先，结果表明BMF是miR-640的靶基因。miR-640在GBM细胞中负向调节BMF的表达。此外，研究结果显示，抑制miR-640可通过靶向BMF显著抑制U251细胞增殖、促进细胞凋亡并提高GBM细胞对TMZ的敏感性。此外，BMF过表达可显著抑制U251细胞增殖、诱导细胞凋亡并提高GBM细胞对TMZ的敏感性。抑制miR-640表达可通过靶向BMF增强人GBM细胞对TMZ的化疗敏感性。
