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KMID : 1034820210170010069
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Molecular & Cellular Toxicology
2021 Volume.17 No. 1 p.69 ~ p.78
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MiR-193b enhanced proliferation and migration and inhibits apoptosis through targeting RAB7A in osteosarcoma cell
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Zhang Yuan-Yuan
Xu Hai-Yan
Dai Jing-Jing
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Abstract
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Background: Accumulating study indicated that microRNA (miRNA) played critical role in the osteosarcoma (OS). The role and mechanisms of miR-193b in OS cell lines were still unknown.
Objective: We resolved the miR-193b expression in OS cell line and normal cell by RT-PCR assay. The effects of upregulated miR-193b on OS cell proliferation, migration, invasion and apoptosis were evaluated using CCK-8 assay, transwell assay, would-healing assay and flow cytometric analysis in vitro, respectively. We investigated the effect of upregulated miR-193b on the mRNA level of cell cycle protein CCND1 and CCNE1 using RT-PCR assay and the protein level of epithelial to mesenchymal transition (EMT)-related protein E-cadherin, vimentin, and N-cadherin by western blotting assay in MG-63 and U2SO cells. Furthermore, luciferase reporter assays were employed to identify the candidate target gene RAB7A of miR-193b.
Results: The expression of miR-193b was downregulated in OS cells. In MG-63 and U2SO cells, ectopic miR-193b expression inhibited cell proliferation, migration, invasion, and induced apoptosis. We found that miR-193b reduced the mRNA expression of CCND1 and CCNE1, and regulated the associated proteins of EMT including E-cadherin, vimentin and N-cadherin in MG-63and U2SO cell lines. Moreover, the candidate target gene RAB7A was negatively regulated by miR-193b. In addition, upregulated RAB7A rescued the inhibitory effect of miR-193b mimics on the development of OS cell.
Conclusion: In conclusion, this study suggested that miR-193b overexpression inhibited cell proliferation, migration, invasion, and induced cell apoptosis by down-regulating RAB7A in OS cell lines.
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KEYWORD
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MiR-193b, Osteosarcoma, RAB7A, EMT
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