LncRNA SNHG9 Promotes Osteosarcoma Progression Through Regulating MiR-1286/HK2

Objective:Long non-coding RNA(Lnc RNA)plays an important role in regulating the development of tumors.Lnc RNA SNHG9 is known to function as an oncogene and to be overexpressed in a variety of cancers.But the precise mechanisms by which Lnc RNA SNHG9 causes osteosarcoma to become malignant are still unknown.The goal of this research was to look into the biological functions and specific mechan-isms of Lnc RNA SNHG9 in osteosarcoma.Methods:First,we obtained the osteosarcoma microarray(GSE126209)from the online biology library GEO,performed bioinformatics analysis on this osteosarcoma microarray,and predicted the expression level of Lnc RNA SNHG9 in osteosarcoma.Real-time fluorescence quantitative PCR(q RT-PCR)was used to detect the expression of Lnc RNA SNHG9 in several osteosarcoma cell lines.The 143B(knockdown),MG63,U2R(overexpression),and HOS were selected for in vitro functional assays of Lnc RNA SNHG9.The effects of Lnc RNA SNHG9,mi RNA-1286,and HK2 on the proliferation and metastasis of osteosarcoma cells were investigated by in vitro functional assays(including: transwell assay,MTT assay,cell scratch assay,clone formation assay,flow cytometry analysis,etc.).Dual luciferase reporter,q RT-PCR,and protein immunoblotting(Western blot)were used to verify whether there was an interaction between Lnc RNA SNHG9 and mi RNA-1286.Lnc RNA SNHG9 was overexpressed or knocked down in osteos-arcoma cells,and overexpression or knockdown efficiency was detected by q RT-PCR.We used the MTT assay to detect tumor cell proliferation,the transwell assay to detect cell migration and invasion,and flow cytometry analysis to identify apoptosis.Western blot was used to detect the expression of apoptosis,EMT-related proteins,and the downstream protein HK2.Lastly,we used an in vivo nude mouse tumorigenesis assay to determine whether Lnc RNA SNHG9 regulates osteosarcoma growth in organisms.Results:Through bioinformatics analysis,we found that Lnc RNA SNHG9 was signif-icantly more expressed in osteosarcoma tissues than in osteosarcoma paracancer tissues in this study.Furthermore,q RT-PCR was used to confirm the Lnc RNA SNHG9’s upregulation in osteosarcoma cell lines.Osteosarcoma cells’ capacity to proliferate,migrate,and invade was decreased,and apoptosis was encouraged by Lnc RNA SNHG9 knockdown.In contrast,high levels of Lnc RNA SNHG9 expression had the opposite effect.Lnc RNA SNHG9 functions as a sponge for mi RNA-1286,regulating the HK2 signaling pathway.Moreover,the addition of mi R-588 mimics or inhibitors reversed the effects of overexpressing or silencing SNHG9,respectively.Finally,in the tumorigenic experiment in nude mice,the volume and mass of osteosarcoma were significantly lower in the knockdown Lnc RNA SNHG9 group than in the control group.Conclusion:In this study,we investigated that Lnc RNA SNHG9 could regulate apoptosis,proliferation and metastasis of osteosarcoma cells through mi RNA-1286/HK2 axis,and preliminary validation was obtained in mice.This may provide a new therapeutic target for the future treatment of osteosarcoma.