神经胶质瘤新癌靶点的发现具有重要意义。我们在这里探讨了神经胶质瘤中与 ORC6（起源识别复合物 6）相关的表达模式、生物学功能和潜在机制。通过生物信息学分析，我们发现人胶质瘤组织中 ORC6 表达显着增加，与较差的总体生存率、较高的肿瘤级别和野生型异柠檬酸脱氢酶状态相关。此外，在从局部治疗的患者获得的神经胶质瘤组织以及各种原代/已建立的神经胶质瘤细胞中检测到 ORC6 过度表达。进一步的生物信息学检查表明，与 ORC6 共表达的基因在与癌症相关的多个信号级联中富集。在原代和永生化 (A172) 神经胶质瘤细胞中，使用特定 shRNA 或 Cas9-sgRNA 敲除 (KO) 来耗尽 ORC6 显着降低细胞活力和增殖，破坏细胞周期进程和流动性，并引发细胞凋亡。相反，通过慢病毒构建体增强 ORC6 表达会增强人神经胶质瘤细胞的恶性行为。 ORC6 成为神经胶质瘤细胞内关键致癌基因（包括 Cyclin A2、Cyclin B2 和 DNA 拓扑异构酶 II (TOP2A)）表达的关键调节因子。 ORC6的沉默或敲除降低了这些基因的mRNA和蛋白质水平，而ORC6的过度表达则增加了它们在原代神经胶质瘤细胞中的表达。生物信息学分析进一步确定RBPJ是ORC6的潜在转录因子。 RBPJ shRNA 降低了原代胶质瘤细胞中 ORC6 的表达，而其过度表达则增加了 ORC6 的表达。此外，在神经胶质瘤组织和细胞中检测到 RBPJ 蛋白与拟议的 ORC6 启动子区域之间的结合显着增强。体内实验表明，ORC6 KO 后，小鼠大脑中患者来源的神经胶质瘤异种移植物的生长显着减少。在这些 ORC6 KO 颅内神经胶质瘤异种移植物中检测到 ORC6 耗竭、增殖抑制、Cyclin A2/B2/TOP2A 表达减少以及细胞凋亡增加。总而言之，RBPJ 驱动的 ORC6 过度表达可促进神经胶质瘤细胞生长，强调其作为有前景的治疗靶点的重要性。© 2024。作者。

The discovery of novel oncotargets for glioma is of immense significance. We here explored the expression patterns, biological functions, and underlying mechanisms associated with ORC6 (origin recognition complex 6) in glioma. Through the bioinformatics analyses, we found a significant increase in ORC6 expression within human glioma tissues, correlating with poorer overall survival, higher tumor grade, and wild-type isocitrate dehydrogenase status. Additionally, ORC6 overexpression is detected in glioma tissues obtained from locally-treated patients and across various primary/established glioma cells. Further bioinformatics scrutiny revealed that genes co-expressed with ORC6 are enriched in multiple signaling cascades linked to cancer. In primary and immortalized (A172) glioma cells, depleting ORC6 using specific shRNA or Cas9-sgRNA knockout (KO) significantly decreased cell viability and proliferation, disrupted cell cycle progression and mobility, and triggered apoptosis. Conversely, enhancing ORC6 expression via a lentiviral construct augmented malignant behaviors in human glioma cells. ORC6 emerged as a crucial regulator for the expression of key oncogenic genes, including Cyclin A2, Cyclin B2, and DNA topoisomerase II (TOP2A), within glioma cells. Silencing or KO of ORC6 reduced the mRNA and protein levels of these genes, while overexpression of ORC6 increased their expression in primary glioma cells. Bioinformatics analyses further identified RBPJ as a potential transcription factor of ORC6. RBPJ shRNA decreased ORC6 expression in primary glioma cells, while its overexpression increased it. Additionally, significantly enhanced binding between the RBPJ protein and the proposed ORC6 promoter region was detected in glioma tissues and cells. In vivo experiments demonstrated a significant reduction in the growth of patient-derived glioma xenografts in the mouse brain subsequent to ORC6 KO. ORC6 depletion, inhibited proliferation, decreased expression of Cyclin A2/B2/TOP2A, and increased apoptosis were detected within these ORC6 KO intracranial glioma xenografts. Altogether, RBPJ-driven ORC6 overexpression promotes glioma cell growth, underscoring its significance as a promising therapeutic target.© 2024. The Author(s).