Circ＿0000518 Competitively Combined With MiR-326 To Regulate SLC27A4 On Invasion And Migration Of Synovial Sarcoma

Objective: A preliminary study was conducted to investigate the differential circ＿0000518 binding to miR-326 as a competitive endogenous RNA(ce RNAs),thereby regulating the expression of the target gene SLC27 A and affecting the proliferation,invasion and migration ability of SS.Methods:(1)The expression of circ＿0000518 in SS tissues was detected by q RT-PCR.(2)Using bioinformatics analysis,the downstream miRNAs and m RNA of circ＿0000518 were obtained through database retrieval and intersection.(3)The primer sequences of miR-326 and SLC27A4 were designed and the expression of miR-326 and SLC27A4 was detected by q RT-PCR in synovial sarcoma tissue,circ＿0000518 overexpressed and knockout cells.(4)SYT-SSX1 overexpressed stable cell lines were constructed in SW982 cell lines using the lentvirus vector SYT-SSX1,and the transfection efficiency of SYT-SSX1 overexpressed stable cell lines was detected by RT-PCR.(5)SW982 cell line was transfected with hsa＿circ＿0000518 overexpression and knockdown plasmid by transient plasmid transfection,and the transfection efficiency was detected by q RT-PCR.(6)The changes of proliferation,invasion and migration of circ＿0000518 silenced and overexpressed SW982 cells were detected by CCK-8,scratch assay and Transwell assay.(7)The Spearman correlation analysis,normality test and T-test were all statistically analyzed using the SPSS26.0 software.?Results:(1)circ＿0000518 was overexpressed in SS and closely correlated with FNCLCC grade and distant metastasis.(2)circ RNAs and miRNA related databases showed that miR-326 and SLC27A4 were the downstream miRNA and m RNA of circ＿0000518,and their binding sites were predicted.(3)q RT-PCR results showed that the expression of miR-326 is low in SS tissues and is negatively correlated with circ＿0000518.SLC27 A was overexpressed in SS,and is positively correlated with circ＿0000518,while negatively correlated with miR-326.(4)The clinicopathological analysis of miR-326,SLC27A4 and SS patients found that the expression of miR-326 and SLC27A4 was closely related to the TNM staging of SS.(5)In the overexpressed circ＿0000518 cells,the expression of miR-326 was increased while the expression of SLC27A4 was decreased;on the other hand,after the knockout of circ＿0000518,the expression of SLC27A4 was increased and the expression of miR-326 was decreased,and SYT-SSX1 type fusion gene overexpressed polyclonal stable cell lines were constructed successfully.(6)After circ＿0000518 was overexpressed in SW982 cells and SYT-SSX1 cells,the proliferation,migration and invasion of circ＿0000518 were significantly enhanced.(7)After knockdown circ＿0000518,the proliferation,migration and invasion ability of SW982 cell line and SYT-SSX1 cells were significantly inhibited.Conclusions: 1.miR-326 and SLC27A4 are downstream miRNAs and m RNA of circ＿0000518;2.High expression of circ＿0000518 in SS was negatively correlated with low expression of miR-326 and positively correlated with high expression of SLC27A4;3.Overexpression of circ＿0000518 enhances SS cell proliferation,migration and invasion,inhibition of miR-326 and enhancement of SLC27A4,and inhibition of circ＿0000518 inhibits SS cell proliferation.Migration and invasion capacity and inhibition of miR-326 are released,which reduces the expression of SLC27A4.?...