Department of Pediatric Surgery, the Affiliated Hospital of Qingdao University, Qingdao, Shandong, China.
Urology, the Affiliated Hospital of Qingdao University, Qingdao, Shandong, China.
Bioengineered. 2020 Dec;11(1):219-228. doi: 10.1080/21655979.2020.1729928.
Neuroblastoma (NB) is responsible for 15% of all childhood cancer deaths. Despite advances in treatment and disease management, the overall 5-year survival rates remain poor in high-risk disease (25-40%). It is well known that miR-145 functions as a tumor suppressor in several types of cancer. However, the impact of miR-145 on NB is still ambiguous. Our aim was to investigate the potential tumor suppressive role and mechanisms of miR-145 in high-risk neuroblastoma. Expression levels of miR-145 in tissues and cells were determined using RT-qPCR. The effect of miR-145 on cell viability was evaluated using MTT assays, apoptosis levels were determined using TUNEL staining, and the MTDH protein expression was determined using western blot and RT-PCR. Luciferase reporter plasmids were constructed to confirm direct targeting for MTDH. that miR-145 expression was significantly lower in high-risk MYCN amplified (MNA) tumors and low miR-145 expression was associated with worse EFS and OS in our cohort. Over-expression of miR-145 reduced cell viability and increased apoptosis in SH-SY-5Y cells. We identified MTDH as a direct target for miR-145 in SH-SY-5Y cells. Targeting MTDH has the similar results as miR-145 overexpression. Our findings suggest that low miR-145 expression was associated with poor prognosis in patients with NB, and the overexpression of miR-145 inhibited NB cells growth by down-regulating MTDH, thus providing a potential target for the development of microRNA-based approach for NB therapy.
神经母细胞瘤（NB）是导致所有儿童癌症死亡的 15%。尽管在治疗和疾病管理方面取得了进展，但高危疾病（25-40%）的总体 5 年生存率仍然较差。众所周知，miR-145 在几种类型的癌症中起肿瘤抑制作用。然而，miR-145 对 NB 的影响仍然不清楚。我们的目的是研究 miR-145 在高危神经母细胞瘤中的潜在肿瘤抑制作用和机制。使用 RT-qPCR 确定组织和细胞中 miR-145 的表达水平。使用 MTT 测定法评估 miR-145 对细胞活力的影响，使用 TUNEL 染色测定细胞凋亡水平，使用 Western blot 和 RT-PCR 测定 MTDH 蛋白表达。构建荧光素酶报告质粒以确认对 MTDH 的直接靶向作用。发现高危 MYCN 扩增（MNA）肿瘤中 miR-145 的表达明显降低，我们的队列中低 miR-145 表达与 EFS 和 OS 较差相关。miR-145 的过表达降低了 SH-SY-5Y 细胞的活力并增加了细胞凋亡。我们确定 MTDH 是 SH-SY-5Y 细胞中 miR-145 的直接靶标。靶向 MTDH 具有与 miR-145 过表达相似的结果。我们的研究结果表明，NB 患者中低 miR-145 表达与预后不良相关，miR-145 的过表达通过下调 MTDH 抑制 NB 细胞生长，从而为 miRNA 为基础的 NB 治疗方法的开发提供了潜在的靶标。
