Interference of ROR1 gene expression inhibits epithelial-mesenchymal transition of osteosarcoma MG-63 cells
10.3781/j.issn.1000-7431.2018.11.743
- Author:
Xingwen WANG
1
Author Information
1. Department of Orthopedics, Second Hospital of Lanzhou University
- Publication Type:Journal Article
- Keywords:
Epithelial-mesenchymal transition;
Osteosarcoma;
Receptor tyrosine kinase-like orphan receptor 1;
RNA interference
- From:
Tumor
2018;38(3):173-181
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To investigate the effect of interferencing receptor tyrosine kinase-like orphan receptor 1 (ROR1) gene expression on epithelialmesenchymal transition (EMT) of osteosarcoma MG-63 cells. Methods: The specific siRNA targeting ROR1 gene (ROR1-siRNA) was transfected into osteosarcoma MG-63 cells with the relative high expression of ROR1 protein, then the down-regulation of ROR1 expression was confirmed by Western blotting. The migration and invasion abilities of MG-63 cells were examined by scratch wound healing assay and Transwell chamber assay. The morphological changes of MG-63 cells were observed under an inverted optical microscope. Finally, the expressions of EMT-associated E-cadherin and vimentin, and tumor metastasis-related zinc finger E-box binding homeobox protein 1 (ZEB1), matrix metalloproteinase 2 (MMP-2) and MMP-9 proteins in MG-63 cells were detected by Western blotting and immunofluoresence staining, respectively. Results: ROR1-siRNA transfection significantly inhibited the expression of ROR1 in MG-63 cells (P < 0.05). After down-regulating ROR1 gene expression by ROR1-siRNA transfection, the migration and invasion abilities of MG-63 cells were significantly reduced (both P < 0.05), the morphology of MG-63 cells conversed from mesenchymal phenotype to epithelial phenotype, and the expression level of E-cadherin was significantly up-regulated (P < 0.05), while the expression level of vimentin was significantly down-regulated (P < 0.05). Furthermore, the expressions of ZEB1, MMP-2 and MMP-9 in MG-63 cells transfected with ROR1-siRNA were decreased as compared with those in the untransfected group (all P < 0.05). Conclusion: RNA interference of ROR1 gene expression can reduce the migration and invasion abilities of osteosarcoma MG-63 cells through inhibiting the occurrence of EMT.
