Bu Jie, Guo Ru, Xu Xue-Zheng, Luo Yi, Liu Jian-Fan
Department of Orthopaedics, Hunan Cancer Hospital and The Affiliated Cancer Hospital of Xiangya School of Medicine, Central South University, Changsha 410013, Hunan Province, People's Republic of China.
Department of Pediatrics, Maternal and Child Health Care Hospital of Hunan Province, Changsha 410008, Hunan Province, People's Republic of China.
J Bone Oncol. 2021 Jan 19;27:100348. doi: 10.1016/j.jbo.2021.100348. eCollection 2021 Apr.
Osteosarcoma is a primary cause of cancer-associated death in children and adolescents worldwide. Long non-coding RNAs SNHG16 (lncRNA SNHG16) and integrin subunit-a 6 (ITGA6) are recently reported to be involved in the tumorigenesis of osteosarcoma by multiple mechanisms. However, the correlation between SNHG16 and ITGA6 in osteosarcoma remains undetermined.
Expression of miR-488, SNHG16 and ITGA6, as well as epithelial-mesenchymal transition (EMT) associated markers in osteosarcoma tissues and cell lines were examined by qRT-PCR or Western blotting. Effects of miR-488, SNHG16 and ITGA6 on cell migration, invasion were evaluated by wound-healing assay and transwell assay. Bioinformatics analysis and dual-luciferase reported assays were applied to assess the interaction among miR-488, SNHG16 and ITGA6. RNA immunoprecipitation (RIP) was also used to verify SNHG16 and miR-488 interaction. Finally, animal study was used to detect the effect of SNHG16 on osteosarcoma .
SNHG16 and ITGA6 were significantly increased while miR-488 was decreased in osteosarcoma. ITGA6 was screened as a target gene of miR-488, and SNHG16 was sponged by miR-488 in osteosarcoma cells. MiR-488 overexpression and SNHG16 knockdown suppressed migration, invasion and EMT of osteosarcoma cells. Moreover, rescue assays proved that the influences of SNHG16 on osteosarcoma cells migration, invasion and EMT were dependent on miR-488 and ITGA6. In addition, the promotive effects of SNHG16 on osteosarcoma tumor growth and metastasis were further supported by xenograft tumor growth assay.
SNHG16 promoted migration, invasion and EMT of osteosarcoma by sponging miR-488 to release ITGA6.
骨肉瘤是全球儿童和青少年癌症相关死亡的主要原因。长链非编码RNA SNHG16（lncRNA SNHG16）和整合素亚基α6（ITGA6）最近被报道通过多种机制参与骨肉瘤的肿瘤发生。然而，骨肉瘤中SNHG16与ITGA6之间的相关性仍未确定。
通过qRT-PCR或蛋白质免疫印迹法检测骨肉瘤组织和细胞系中miR-488、SNHG16和ITGA6的表达，以及上皮-间质转化（EMT）相关标志物。通过伤口愈合试验和Transwell试验评估miR-488、SNHG16和ITGA6对细胞迁移、侵袭的影响。应用生物信息学分析和双荧光素酶报告基因试验评估miR-488、SNHG16和ITGA6之间的相互作用。RNA免疫沉淀（RIP）也用于验证SNHG16与miR-488的相互作用。最后，通过动物研究检测SNHG16对骨肉瘤的影响。
骨肉瘤中SNHG16和ITGA6显著升高，而miR-488降低。ITGA6被筛选为miR-488的靶基因，在骨肉瘤细胞中SNHG16被miR-488吸附。miR-488过表达和SNHG16敲低抑制了骨肉瘤细胞的迁移、侵袭和EMT。此外，挽救试验证明SNHG16对骨肉瘤细胞迁移、侵袭和EMT的影响依赖于miR-488和ITGA6。此外，异种移植瘤生长试验进一步支持了SNHG16对骨肉瘤肿瘤生长和转移的促进作用。
SNHG16通过吸附miR-488释放ITGA6促进骨肉瘤的迁移、侵袭和EMT。
