Investigation On The Mechanism Of Propofol Attenuates Cell Progression Of Osteosarcoma By Up-regulating LncRNA TUSC7 Expression And Inactivating AKT/GSK3β Signaling

BackgroundPropofol is a kind of intravenous anesthetic,which belongs to one of the commonly used anesthetics in clinic.In recent years,the anti-cancer effect of propofol has attracted the attention of clinical and basic researchers,and the use of propofol in intraoperative anesthesia and postoperative analgesia plays an important role in the elimination of micrometastases and residual tumor cells after tumor surgery.From the perspective of molecular pathological mechanism,propofol inhibits the activity of tumor cell phenotypes(such as proliferation,invasion,metastasis,apoptosis,etc.)by regulating a variety of signaling pathways,especially its potential regulatory effect on non-coding RNA(ncRNA),which provides a promising direction for exploring new targets for cancer therapy.Osteosarcoma is a primary malignant bone tumor,distributed commonly in children and young adults,but its etiology and pathologic mechanism are still unclear.Further exploration of the underlying molecular mechanisms of tumor development is important to improve diagnosis and treatment for osteosarcoma.LncRNAs in tumor cells have diagnostic and prognostic potential,and can be used as therapeutic targets or markers for cancer prevention and prognosis,providing new opportunities for cancer diagnosis and treatment,which has attracted extensive attention of researchers.In osteosarcoma,the functions of some lncRNAs have been annotated,which can promote or inhibit the occurrence and progression of osteosarcoma.The role of these lncRNAs in propofol against osteosarcoma is unknown.This study aims to explore whether lncRNA plays a role in the inhibition of osteosarcoma phenotype by propofol,and what the mechanism is.An in-depth understanding of the molecular mechanism of lncRNA can not only pave the way for the exploration of the pathogenesis of osteosarcoma,but also provide lncRNA-based therapies to improve patient prognosis and improve patient survival.Research contentObjective:To screen lncRNAs that can be affected by propofol,and pave the way for further exploration of the mechanism of propofol relieving osteosarcoma through lncRNA molecules.Methods:1.Real-time quantitative PCR was used to detect the expression level of pro-cancer or anti-cancer lncRNAs in propofol-treated human osteosarcoma cell line U-2 OS.2.Cell activity,migration and invasion activities were observed in U-2 OS cells with TUSC7 silence(si-TUSC7 transfected into cells)and propofol treatment.3.EMT associated protein expression was detected by Western blot.Results:1.Propofol promoted the expression of anti-tumor lncRNA TUSC7 in osteosarcoma cells.2.Silencing lncRNA TUSC7 attenuated the inhibitory effect of propofol on proliferation,migration and invasion of osteosarcoma cells.3.Silencing lncRNA TUSC7 attenuated the promoting or inhibiting effects of propofol on the expression of EMT-related proteins.Conclusion:Propofol promotes TUSC7 expression,and suppresses cell viability,migration and invasion.Objective:To investigate whether FoxO1 mediates propofol to promote TUSC7 expression and the interaction between FoxO1 and TUSC7.Methods:1.The expression levels of whole cell FoxO1(total-FoxO1),phosphorylated FoxO1(p-FoxO1)and nucleus FoxO1 were detected by Western blot.2.Chromatin Immunoprecipitation(ChIP)was used to detect the FoxO1 binding TUSC7 promoter region.3.Interaction between FoxO1 and TUSC7 promoter was detected by dual luciferase reporter gene.Results:1.Propofol inhibited FoxO1 phosphorylation in osteosarcoma cells,and promoted whole-cell FoxO1 expression and FoxO1 nucleation.2.Simultaneously silencing FoxO1 expression(cell transfection with si-FoxO1)significantly reduced TUSC7 expression compared with propofol treatment.3.FoxO1 overexpression significantly increased TUSC7 expression,while FoxO1 silencing significantly decreased TUSC7 expression.4.FoxO1 binded to the TUSC7 promoter region and positively regulated TUSC7 promoter transcription.5.Overexpression of TUSC7 did not affect FOXO1 expressionConclusion:propofol regulates TUSC7 expression by regulating FoxO1,and FoxO1 interacts with TUSC7.Objective:To investigate the key role of FoxO1/TUSC7 signaling axis in the anti-proliferation,migration and invasion effect of propofol on osteosarcoma cells.Methods:To observe the effect of FoxO1/TUSC7 signaling pathway on the inhibition of tumor growth and metastasis by propofol,U-2 OS cells were divided into four groups:control group,propofol +si-NC+pcDNA-NC group,propofol+si-FoxO1+pcDNA-NC group,and propofol+si-FoxO1+pcDNA-TUSC7 group.Results:1.Silencing of transcription factor FoxO1 attenuated the effect of propofol and significantly increased cell activity,migration and invasion;2.Overexpression of TUSC7 effectively reversed the effect of si-FoxO1 and reduced the activity,migration and invasion of cells.Conclusion:Propofol suppresses cell viability,migration and invasion by regulating FoxO1/TUSC7siganling pathway.Objective:To investigate the key role of AKT/GSK3β signaling pathway in anti-proliferation,migration and invasion effect of propofol on osteosarcoma cells.Methods:1.In order to determine the effect of AKT/GSK3β signaling pathway on the inhibition of tumor cell growth by propofol,U-2 OS cells were divided into four groups:control group,propofol group,propofol+pcDNA-NC group,and propofol +pcDNA-GSK3β group(propofol treated at a concentration of 5μg/mL).The treated cells were collected and detected for cell activity,migration and invasion.2.The expression levels of AKT and its phosphorylation(p-Akt),GSK3β and its phosphorylation(p-GSK3β)were detected by Western blot.Results:1.Propofol decreased the phosphorylation of AKT and GSK3β(p-AKT and p-GSK3β)and inhibited the activation of Akt/GSK3β signaling pathway.2.Overexpression of GSK3β attenuated the effect of propofol and increased proliferation,invasion and migration of osteosarcoma cells.3.TUSC7 silencing attenuated the effect of propofol and increased the phosphorylation of AKT and GSK3β.Conclusion:Propofol promotes the expression of lncRNA TUSC7 by promoting the expression of transcription factor FoxO1 and nucleation,thereby inhibiting the proliferation,migration and invasion of osteosarcoma cells.In addition,propofol also inhibits the proliferation,invasion and migration of osteosarcoma cells by inhibiting the activation of AKT/GSK3β signaling pathway.