Department of Neurosurgery, Second Affiliated Hospital of Nanchang University, No. 1, Minde Road, Nanchang, 330006, Jiangxi Province, People's Republic of China.
J Neurooncol. 2021 Sep;154(2):131-144. doi: 10.1007/s11060-021-03814-7. Epub 2021 Aug 6.
Circular RNA (circRNA) has been demonstrated to play key roles in regulating glioma progression. Understanding the regulatory mechanism of circRNA in glioma is vital to reveal the pathogenesis of glioma and develop novel therapeutic strategies. Therefore, our study focuses on the role and underlying mechanism of Circ_CLIP2 in glioma.
The expression of Circ_CLIP2, miR-195-5p and HMGB3 in glioma cells and tissues were analyzed using qRT-PCR. Cell proliferation was determined with colony formation and MTT assays. Cell cycle and apoptosis were examined by flow cytometry. Western blot was conducted for analyzing HMGB3, PCNA, Bax, Bcl-2, cleaved-caspase 3, Wnt-1 and β-catenin. Dual-luciferase reporter assay was measured to investigate the interaction among Circ_CLIP2, miR-195-5p and HMGB3.
The expression of Circ_CLIP2 and HMGB3 were increased while miR-195-5p was down-regulated in glioma cells and patients. Silencing of Circ_CLIP2 inhibited cell proliferation, enhanced cell apoptosis and inhibited the Wnt/β-catenin signaling pathway. Circ_CLIP2 suppressed miR-195-5p expression by directly sponging miR-195-5p. MiR-195-5p inhibited HMGB3 expression via directly targeting HMGB3. Knockdown of miR-195-5p facilitated cell proliferation, inhibited cell apoptosis and activated Wnt/β-catenin signaling, which were reversed by silencing of HMGB3.
Knockdown of Circ_CLIP2 suppresses glioma progression by targeting miR-195-5p/HMGB3 thus inhibiting Wnt/β-catenin signaling. This study may provide potential therapeutic targets against glioma.
环状 RNA（circRNA）已被证明在调控神经胶质瘤进展中发挥关键作用。了解 circRNA 在神经胶质瘤中的调控机制对于揭示神经胶质瘤的发病机制和开发新的治疗策略至关重要。因此，我们的研究重点是 Circ_CLIP2 在神经胶质瘤中的作用及其潜在机制。
采用 qRT-PCR 分析神经胶质瘤细胞和组织中 Circ_CLIP2、miR-195-5p 和 HMGB3 的表达。采用集落形成和 MTT 检测分析细胞增殖。采用流式细胞术检测细胞周期和凋亡。采用 Western blot 分析 HMGB3、PCNA、Bax、Bcl-2、cleaved-caspase 3、Wnt-1 和 β-catenin。采用双荧光素酶报告基因检测分析 Circ_CLIP2、miR-195-5p 和 HMGB3 之间的相互作用。
Circ_CLIP2 和 HMGB3 的表达在神经胶质瘤细胞和患者中升高，而 miR-195-5p 下调。Circ_CLIP2 沉默抑制细胞增殖，增强细胞凋亡，抑制 Wnt/β-catenin 信号通路。Circ_CLIP2 通过直接海绵吸附 miR-195-5p 抑制 miR-195-5p 的表达。miR-195-5p 通过直接靶向 HMGB3 抑制 HMGB3 的表达。miR-195-5p 的敲低促进细胞增殖，抑制细胞凋亡，激活 Wnt/β-catenin 信号通路，而沉默 HMGB3 则逆转了这些作用。
Circ_CLIP2 通过靶向 miR-195-5p/HMGB3 抑制 Wnt/β-catenin 信号通路抑制神经胶质瘤的进展。本研究可为神经胶质瘤的治疗提供潜在的靶点。
