| Objective: To explore the expression of long non-coding RNASBF2-AS1 in esop hageal squamous cell carcinoma(ESCC),and observe the effect of its mediated ceRNA network mechanism on the proliferation and radiotherapy sensitivity of E SCC.Methods: Detection of the expression level of LncRNASBF2-AS1 in esophageal normal epithelial cells and esophageal squamous cell carcinoma cell lines by RT-PCR.Two cells with high expression in ESCC cells,ECA109 and TE-13,were selected and divided into si-SBF2-AS1 group,si-NC group,SBF2-AS1 and PCD NA group.Through RT-PCR,RTCA,clonal formation,EDU experiments to detec t the proliferation of various groups of cells in ESCC.Flow Cytometry Test to detect the effect of SBF2-AS1 on the cycle of esophageal squamous cellcarcinom a cells.The tumor-bearing experiment in nude mice tested the effect of silencing SBF2-AS1 on the proliferation of ESCC.The effect of SBF2-AS1 on the radios ensitivity of esophageal squamous cell carcinoma was detected by themeasuremen t of radiosensitivity and cell survival test.Two target genes miR-338-3P and miR-362-3P that may be regulated by SBF2-AS1 are predicted by Starbase V2.0.RTPCR was used to detect the expression levels of miR-338-3P and miR-362-2P in ESCC cell lines.The binding between SBF2-AS1 and miR-338-3P andmiR-362-3Pwas verified by RIP and pulldown experiments.The luciferase reporter gene and pulldown experiment verified that miR-338-3P and miR-362-3P canbind to E2F1.The rescue experiment and westernblot experiment further verified the relationshi p between SBF2-AS1,miR-338-3P,miR-362-3P and E2F1.And through rescue e xperiments to verify the effects of SBF2-AS1/miR-338-3P,miR-362-3P/E2F1 axis on the proliferation and radiosensitivity of esophageal squamous cellcarcinoma.Results: SBF2-AS1 is highly expressed in ESCC tissues and cell lines(P<0.05).Inhibition of SBF2-AS1 can inhibit theproliferation of ESCC(P<0.05),and overe xpression of SBF2-AS1 can promote the proliferation of ESCC(P<0.05).SBF2-AS1 promotes the proliferation of ESCC cells by promoting the transformation o f G1 into S phase(P<0.05).In vivo experiments further confirmed that the tumo r size that interfered with SBF2-AS1 group was significantly smaller than that of NC group(P<0.05).Under the same dose of radiation,radiosensitivity measurem ent and cell survival experiments show that overexpression of SBF2-AS1 can red uce the radiosensitivity of esophageal squamous cell carcinoma,while interference with SBF2-AS1 can increase the radiosensitivity of esophageal squamous cell,co mpared with the NC group(P<0.05).Mechanism studies confirmed that SBF2-A S1 can be used as ce RNA to bind miR-338-3P and miR-362-3P(P<0.05).The p ullddown experimentof luciferase reporter gene showed that miR-338-3P and miR-362-3P can bind to E2F1(P<0.05).Rescue experiments andwestern blot experim ents show that SBF2-AS1 regulates E2F1 expression bycompetitively binding miR-338-3p and miR-362-3p,and miR-338-3P and miR-362-3P can alleviate the eff ects of SBF2-AS1 on the esophagus to a certain extent(P<0.05).The last rescue experiment showed that SBF2-AS1 promotes the proliferation of esophageal squa mous cell carcinoma and increases its radiation resistance through miR-338-3P an d miR-362-3P(P<0.05).Conclusion: SBF-AS1 plays a role as an oncogene in ESCC,and SBF2-AS1 m ay be one of the reasons for the resistance of ESCC to radiotherapy.Interfering with SBF2-AS1 increases the radiosensitivity of ESCC.The underlying mechanism may be as a ceRNA competitively combining miR-338-3P and miR-362-3P to up-regulate the expression of E2F1,promote the conversion of G1 phase to S phase,so as to promote the proliferation of ESCC andenhance the radiation sensitivity of ESCC. |
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