ABSTRACT

Glioblastoma multiforme (GBM) is associated with a poor prognosis, and effective treatments are lacking. Our previous studies have shown that miRNA-451 is closely related to the development and progression of glioma. miRNA-451 is a tumor suppressor whose expression is negatively correlated with the WHO grades of gliomas, but its specific mechanism is still unclear. Research shows that NF-κB is highly expressed in early malignant glioma, and thus, the NF-κB signaling pathway has become an important target for the treatment of malignant glioma. Activation of IKK is a critical step in the activation of the classical NF-κB pathway. By performing a bioinformatics analysis, we found that IKKβ is a potential direct target of miRNA-451 in glioma. In this study, we transfected lentivirus expressing miRNA-451 to test the effect of miRNA-451 overexpression on malignant glioma cell lines and confirmed that IKKβ is a target gene of miRNA-451 by luciferase assay. By targeting IKKβ, MTT, cell invasion and wound-healing assays showed that cell proliferation, cell invasion and migration were significantly suppressed in the LV-miRNA-451 group. Western blotting results showed that the expression levels of IKKβ, p-p65, MMP-2, MMP-9, Cyclin D1, p16 and PCNA were significantly decreased in the LV-miRNA-451 group. In vivo, miRNA-451 significantly decreased glioma cell growth, and the survival of BALB/c-A nude mice was significantly prolonged. Immunohistochemistry showed that p-p65, Cyclin D1 and Ki67 expression was significantly reduced in the LV-miRNA-451 group. Taken together, these results suggest that miRNA-451 could regulate the NF-κB signaling pathway by targeting IKKβ, which inhibits glioma cell growth in vitro and in vivo. Therefore, this study may provide novel insight into miRNA-451-targeted therapy for glioma.

摘要

多形性胶质母细胞瘤（GBM）与预后不良有关，并且缺乏有效的治疗方法。我们以往的研究表明，miRNA-451与胶质瘤的发生发展密切相关。miRNA-451是一种肿瘤抑制因子，其表达与胶质瘤的WHO分级呈负相关，但其具体机​​制尚不清楚。研究表明，NF-κB在早期恶性胶质瘤中高表达，因此NF-κB信号通路已成为恶性胶质瘤治疗的重要靶点。IKK 的激活是激活经典 NF-κB 通路的关键步骤。通过进行生物信息学分析，我们发现 IKKβ 是 miRNA-451 在胶质瘤中的潜在直接靶标。在这项研究中，我们转染表达miRNA-451的慢病毒以测试miRNA-451过表达对恶性胶质瘤细胞系的影响，并通过荧光素酶测定证实IKKβ是miRNA-451的靶基因。通过靶向 IKKβ、MTT、细胞侵袭和伤口愈合试验表明，LV-miRNA-451 组的细胞增殖、细胞侵袭和迁移受到显着抑制。Western blotting结果显示LV-miRNA-451组IKKβ、p-p65、MMP-2、MMP-9、Cyclin D1、p16和PCNA的表达水平显着降低。在体内，miRNA-451 显着降低了胶质瘤细胞的生长，显着延长了 BALB/cA 裸鼠的存活时间。免疫组织化学显示，LV-miRNA-451 组中 p-p65、Cyclin D1 和 Ki67 的表达显着降低。综合起来，这些结果表明，miRNA-451 可以通过靶向 IKKβ 来调节 NF-κB 信号通路，从而在体外和体内抑制胶质瘤细胞的生长。因此，这项研究可能为 miRNA-451 靶向治疗胶质瘤提供新的见解。