LncRNA FBXl19-ASL As CeRNA Regulates MiR-346 To Promote Proliferation,invasion And Metastasis Of Osteosarcoma

Part Ⅰ: Expression of FBXL19-AS1 in osteosarcoma tissues and cellsObjective: To explore the expression of long non-coding RNA(Lnc RNA)FBXL19-AS1 in osteosarcoma tissues and cells.Methods: The expression levels of FBXL19-AS1 in osteosarcoma tissues and paraneoplastic tissues were compared by real-time quantitative PCR(q RT-PCR)and osteosarcoma cells(MG63,U2 OS,SAOS2,HOS,143B)and human osteoblasts(h FOB1.19)Expression level comparison.Results: 1.The upregulation of Lnc RNA FBXL19-AS1 was observed in osteosarcoma tissue relative to tumor adjacent tissue.2.The expression of Lnc RNA FBXL19-AS1 in human osteoblasts was up-regulated compared to human osteoblasts.Conclusion:It suggests that FBXL19-AS1 may have biological significance in osteosarcoma.Part Ⅱ: Effect of FBXL19-AS1 on proliferation of osteosarcoma cellsObjective:To investigate the effect of FBXL19-AS1 on the proliferation of MG63 and 143 B osteosarcoma cells.Methods: Lentivirus infection down-regulated FBXL19-AS1 of MG63 and 143 B,CCK-8 detected the inhibitory effect of FBXL19-AS1 on proliferation of osteosarcoma cells MG63 and 143 B,and flow cytometry measured FBXL19-AS1 on cell cycle of osteosarcoma cells MG63 and 143 B.The effect of FBXL19-AS1 on the apoptosis of osteosarcoma cell line MG63 and 143 B was detected by Western blot.The effect of FBXL19-AS1 on the proliferation of osteosarcoma cells in nude mice was detected by subcutaneous tumor xenografts in nude mice.Results:1.The cell proliferation ability of FBXL19-AS1 cells treated with CCK-8 was lower than that of the negative control group.2.The cell cycle of MG63,143 B of FBXL19-AS1 osteosarcoma cell was down-regulated in G1 phase by flow cytometry.3.Western blot analysis showed that down-regulation of FBXL19-AS1 inhibited G1 phase-associated cyclin D1,CDK4 and CDK6 and promoted cyclin-dependent kinase inhibitor P27.4.Subcutaneous xenografts in nude mice down-regulated FBXL19-AS1 osteosarcoma cell transplantation The tumor was reduced compared to the negative control.Conclusion: FBXL19-AS1 promotes the proliferation of osteosarcoma and down-regulates the proliferation of osteosarcoma cells after FBXL19-AS1.Part Ⅲ: Effect of FBXL19-AS1 on the invasion and metastasis of osteosarcoma cellsObjective:To verify the effect of FBXL19-AS1 on invasion and metastasis of osteosarcoma cells MG63 and 143 B.Methods: The effect of FBXL19-AS1 on the migration of osteosarcoma cells was evaluated by cell scratch assay.The effect of FBXL19-AS1 on the invasion and migration of osteosarcoma cells was detected by Transwell assay.Results:1.The down-regulation of FBXL19-AS1 in cell scratch test can decrease the migration ability of osteosarcoma cells;2.Transwell migration assay confirmed that the down-regulation of FBXL19-AS1 in osteosarcoma cells MG63 and 143 B was lower than that in the negative control group;3.Transwell Invasion experiments confirmed that down-regulation of FBXL19-AS1 osteosarcoma cells MG63,143 B than the negative control group cell invasion decreased.Conclusion:FBXL19-AS1 promotes the migration and invasion of osteosarcoma and down-regulates the migration and invasion of osteosarcoma cells after FBXL19-AS1.Part Ⅳ: FBXL19-AS1 as a ce RNA Regulator mi R-346Objective: To verify that FBXL19-AS1 regulates mi R-346 mechanism as ce RNA.Methods: Fluorescence in situ hybridization(FISH)and cytoplasmic nucleus isolation assays were used to detect the localization of FBXL19-AS1 in cells.q-PCR assay was used to detect the expression of mi R-346 in osteosarcoma cells.Combination of FBXL19-AS1 and mi R-346.Results:1.Fluorescence in situ hybridization and cytoplasmic nucleus separation assays detected FBXL19-AS1 mainly in cytoplasm;2.q-PCR assay showed low expression in osteosarcoma cells and showed a significant negative correlation with FBXL19-AS1;3.The luciferin reporter assay experimentally verified direct binding of FBXL19-AS1 to mi R-346.Conclusion: FBXL19-AS1 regulates mi R-346 as a ce RNA.Part Ⅴ: FBXL19-AS1 regulates mi R-346 proliferation,invasion and metastasis of osteosarcoma cellsObjective: To verify the effect of FBXL19-AS1 on mi R-346 proliferation,invasion and metastasis of osteosarcoma cells.Methods: CCK-8 and plate cloning experiments were used to evaluate the effect of FBXL19-AS1 regulating mi R-346 on the proliferation of osteosarcoma cells.Cell scratch experiments were performed to verify the effect of FBXL19-AS1 on the migration of osteosarcoma cells.Transwell assay was used to detect FBXL19.-AS1 regulates the effect of mi R-346 on invasion and migration of osteosarcoma cells.Results:1.CCK-8 experiments verify that FBXL19-AS1 promotes the proliferation of osteosarcoma cells through regulating mi R-346;2.Plate cloning experiments verify that FBXL19-AS1 promotes colony formation of osteosarcoma cells through regulating mi R-346;3.The cell scratch test FBXL19-AS1 promotes the migration of osteosarcoma cells by regulating mi R-346;4.Transwell invasion assays verify that FBXL19-AS1 promotes migration and invasion of osteosarcoma cells by regulating mi R-346.Conclusion: FBXL19-AS1 regulates mi R-346 to promote the proliferation,invasion and metastasis of osteosarcoma cells.