Department of Orthopedics, Wuhan Puren Hospital, Puren Hospital Affiliated of Wuhan University of Science and Technology, Wuhan, Hubei, 430080, Chinam.
Department of Orthopedics, Wuhan Puren Hospital, Puren Hospital Affiliated of Wuhan University of Science and Technology, Wuhan, Hubei, 430080, China.
Acta Biochim Pol. 2023 Apr 23;70(2):313-323. doi: 10.18388/abp.2020_6551.
Osteosarcoma is a highly prevalent type of primary bone tissues in children and young adolescents. Micro-RNA (miR) dysregulation has been linked to osteosarcoma tumorigenesis. The role of miR-196-5p was investigated in modulating the growth and metastatic behaviour of human osteosarcoma cells, along with exploring its mechanism of action. As shown by RT-qPCR expression analysis, osteosarcoma cell lines exhibited prominent (P<0.05) transcriptional repression of miR-196-5p. The latter was thus transiently overexpressed in osteosarcoma cells, which resulted in the loss of cell viability and colony formation via induction of autophagy. The western blot analysis of the autophagy marker proteins revealed that the expression of Beclin 1 and LC3B II proteins was induced by miR-196-5p, whereas that of p62 and LC3BI was repressed. Moreover, osteosarcoma cells overexpressing miR-196-5p showed significantly (P<0.05) lower migration and invasion concerning the control osteosarcoma cells. According to the results of the in-silico analysis, Derlin-1 participates in the regulation of miR-196-5p in osteosarcoma, and this prediction has been validated using a dual luciferase assay. The results indicated that miR-196-5p exerted its molecular role by targeting Derlin-1 at the post-transcriptional level. Summing up, the study revealed the modulatory potential of miR-196-5p/Derlin-1 on osteosarcoma cells and provided insights into the possible implications for the treatment and prognosis of the disease.
骨肉瘤是儿童和青少年中一种高发的原发性骨组织肿瘤。微小 RNA（miRNA）失调与骨肉瘤的肿瘤发生有关。本研究旨在探讨 miR-196-5p 在调节人骨肉瘤细胞生长和转移行为中的作用及其作用机制。RT-qPCR 表达分析显示，骨肉瘤细胞系中 miR-196-5p 的转录抑制明显（P<0.05）。因此，通过瞬时过表达 miR-196-5p，导致细胞活力丧失和集落形成减少，通过诱导自噬。自噬标记蛋白的 Western blot 分析显示，miR-196-5p 诱导 Beclin 1 和 LC3B II 蛋白的表达，而 p62 和 LC3BI 的表达则受到抑制。此外，过表达 miR-196-5p 的骨肉瘤细胞的迁移和侵袭能力明显低于对照组骨肉瘤细胞（P<0.05）。通过计算机分析的结果表明，Derlin-1 参与了骨肉瘤中 miR-196-5p 的调控，这一预测已通过双荧光素酶报告基因实验得到验证。结果表明，miR-196-5p 通过在转录后水平靶向 Derlin-1 发挥其分子作用。综上所述，该研究揭示了 miR-196-5p/Derlin-1 对骨肉瘤细胞的调节潜力，并为该疾病的治疗和预后提供了新的见解。
