Department of Orthopedics, Liaocheng Third People' Hospital, No. 62, Weiyu Road, Liaocheng, 252000, Shandong, China.
Department of Child Protection, Liaocheng Dongchangfu District Maternal and Child Health Hospital, No. 129, Zhenxing West Road, Liaocheng, 252000, Shandong, China.
Mol Biotechnol. 2021 Sep;63(9):807-817. doi: 10.1007/s12033-021-00343-6. Epub 2021 May 26.
Understanding the underlying mechanisms of pediatric osteosarcoma (OS) migration and invasion is important for prognosis and treatment. We tried to measure the expression of long non-coding RNA HLA complex group 18 (HCG18) in OS and reveal its function in the malignant behaviors of OS cells. This study detected the expression of HCG18, miR-188-5p and forkhead box C1 (FOXC1) in OS tissues and cell lines by quantitative real-time PCR (qRT-PCR). The relevance between miR-188-5p and HCG18 or FOXC1 was affirmed by dual-luciferase reporter (DLR) assay. Cell viability was analyzed by MTT assay. Transwell assay was utilized to test cell invasion and migration. FOXC1 protein expression was detected by western blot. HCG18 expression was elevated in OS tissues, and enhanced HCG18 expression was related to metastasis. HCG18 silencing repressed the viability, migration and invasion of OS cells. Moreover, HCG18 interacted with miR-188-5p. MiR-188-5p up-regulation repressed cell viability, invasion and migration in OS cells. FOXC1, a known target of miR-188-5p, was negatively modulated by miR-188-5p. Furthermore, miR-188-5p inhibition or FOXC1 over-expression partially abolished the reduced of cell viability, invasion and migration mediated by HCG18 silencing in OS cell lines. This study revealed that HCG18 knockdown repressed the viability, invasion and migration of OS cells by targeting miR-188-5p and regulating FOXC1 expression. Thus, HCG18/ miR-188-5p/FOX may be a hopeful target for OS therapy.
了解小儿骨肉瘤(OS)迁移和侵袭的潜在机制对于预后和治疗很重要。我们试图测量长非编码 RNA HLA 复合物组 18(HCG18)在 OS 中的表达,并揭示其在 OS 细胞恶性行为中的功能。本研究通过定量实时 PCR(qRT-PCR)检测 OS 组织和细胞系中 HCG18、miR-188-5p 和叉头框 C1(FOXC1)的表达。双荧光素酶报告(DLR)测定证实了 miR-188-5p 与 HCG18 或 FOXC1 之间的相关性。MTT 分析检测细胞活力。Transwell 测定用于测试细胞侵袭和迁移。Western blot 检测 FOXC1 蛋白表达。OS 组织中 HCG18 表达升高,增强的 HCG18 表达与转移有关。HCG18 沉默抑制 OS 细胞的活力、迁移和侵袭。此外,HCG18 与 miR-188-5p 相互作用。miR-188-5p 的上调抑制 OS 细胞的活力、侵袭和迁移。FOXC1 是 miR-188-5p 的已知靶标,受 miR-188-5p 的负调控。此外,miR-188-5p 抑制或 FOXC1 过表达部分消除了 HCG18 沉默在 OS 细胞系中对细胞活力、侵袭和迁移减少的影响。本研究表明,HCG18 敲低通过靶向 miR-188-5p 并调节 FOXC1 表达抑制 OS 细胞的活力、侵袭和迁移。因此,HCG18/miR-188-5p/FOX 可能是 OS 治疗的有希望的靶点。