Medical Department, Qingdao Infectious Disease HospitalQingdao, ShandongP.R. China.
Department of Anesthesia, Qingdao Women and Children's HospitalQingdao, ShandongP.R. China.
Oncol Res. 2020 Feb 7;28(1):95-102. doi: 10.3727/096504019X15742472027401. Epub 2019 Nov 21.
Long noncoding RNA CRNDE (CRNDE) recently emerged as a carcinogenic promoter in various cancers including medulloblastoma. However, the functions and molecular mechanisms of CRNDE to the acquired drug resistance of medulloblastoma are still unclear. The transcript levels of CRNDE were examined in four medulloblastoma cell lines exposed to cisplatin treatment, and IC values were calculated. Effects of CRNDE knockdown or miR-29c-3p overexpression on cell viability, colony formation, apoptosis, migration, and invasion were assessed using the CCK-8, colony formation assay, flow cytometry, and Transwell assays, respectively. RNA pulldown and RNA-binding protein immunoprecipitation (RIP) were performed to confirm the molecular interactions between CRNDE and miR-29c-3p involved in medulloblastoma cells. The in vivo role of CRNDE knockdown or miR-29c-3p overexpression on tumor growth and apoptosis was evaluated in a xenograft mouse model of human medulloblastoma. The transcript levels of lncRNA CRNDE were significantly higher in cisplatin-treated tumor cells with higher IC values. Depletion of CRNDE inhibited tumor cell proliferation and colony formation, induced cell apoptosis, and suppressed migration and invasion in medulloblastoma cells. Moreover, overexpression of miR-29c-3p inhibited tumor cell proliferation and colony formation, migration, and invasion, and enhanced apoptosis and chemosensitivity to cisplatin. In addition, CRNDE was found to act as a miR-29c-3p sponge. Furthermore, in vivo experiments showed the CRNDE/miR-29c-3p interactions involved in medulloblastoma. Our study demonstrates that CRNDE acts as a critical mediator of proliferation, apoptosis, migration, invasion, and resistance to chemotherapeutics via binding to and negatively regulating miR-29c-3p in medulloblastoma cells. These results provide novel molecular targets for treatment of medulloblastoma.
长链非编码 RNA CRNDE（CRNDE）最近在包括髓母细胞瘤在内的各种癌症中被发现是一种致癌促进物。然而，CRNDE 对髓母细胞瘤获得性耐药的功能和分子机制仍不清楚。在四种暴露于顺铂治疗的髓母细胞瘤细胞系中检查了 CRNDE 的转录水平，并计算了 IC 值。使用 CCK-8、集落形成测定、流式细胞术和 Transwell 测定分别评估了 CRNDE 敲低或 miR-29c-3p 过表达对细胞活力、集落形成、细胞凋亡、迁移和侵袭的影响。RNA 下拉和 RNA 结合蛋白免疫沉淀（RIP）实验分别用于证实涉及髓母细胞瘤细胞的 CRNDE 和 miR-29c-3p 之间的分子相互作用。在人髓母细胞瘤异种移植小鼠模型中评估了 CRNDE 敲低或 miR-29c-3p 过表达对肿瘤生长和细胞凋亡的体内作用。在具有更高 IC 值的顺铂处理的肿瘤细胞中，lncRNA CRNDE 的转录水平显着升高。CRNDE 耗竭抑制了髓母细胞瘤细胞的肿瘤细胞增殖和集落形成，诱导细胞凋亡，并抑制迁移和侵袭。此外，miR-29c-3p 的过表达抑制了肿瘤细胞的增殖和集落形成、迁移和侵袭，增强了对顺铂的细胞凋亡和化疗敏感性。此外，CRNDE 被发现作为 miR-29c-3p 的海绵。此外，体内实验显示了 CRNDE/miR-29c-3p 相互作用涉及髓母细胞瘤。我们的研究表明，CRNDE 通过与 miR-29c-3p 结合并负调控髓母细胞瘤细胞中的 miR-29c-3p，作为增殖、凋亡、迁移、侵袭和对化疗药物耐药的关键介质发挥作用。这些结果为治疗髓母细胞瘤提供了新的分子靶标。
