LncGABPB1-AS1 Inhibits The Proliferation And Invasion Of Renal Cell Carcinoma Through MiR-1246/PCK1 Axis

Objective: Kidney cancer accounts for 80% to 90% of kidney malignancies.Renal clear cell carcinoma is the most common type,and it is drug-resistant and nonsensitive to radiotherapy or chemotherapy.Advanced renal clear cell carcinoma is prone to invasion and metastasis,and the prognosis is poor.Therefore,the diagnosis and treatment is a challenge.Understanding the pathogenesis and biological mechanisms of renal clear cell carcinoma may improve diagnosis,treatment,and prognosis.Long chain noncoding RNA(lncRNA)is a RNA molecule with a length of more than 200 nt,and it has mRNA like structure,which usually regulates gene expression in epigenetic regulation and other forms.In recent years,it has become a research hotspot of cancer.Moreover,it has been proved that the effect on the occurrence and development of tumors.LncGABPB1-AS1 is the antisense RNA of GABPB1.It has a inhibiting function for liver cancer,but the mechanism of function is unclear.In order to clarify the role and mechanism of GABPB1-AS1 in renal cell carcinoma,we have carried out relevant research.Materials and methods:1.The lncGABPB1-AS1 expression in fresh frozen renal clear cell carcinoma,adjacent tissue,cultured cancer cell lines and normal ones was detected by RT-PCR.The correlation between the lncGABPB1-AS1 expression and clinicopathological characteristics was analyzed.To construction of lncGABPB1-AS1 vector and transfection of 786-O,Caki-1 cell lines,the proliferation of cancer cells in vivo and in vitro was detected by CCK-8 assays and subcutaneous tumorigenesis in nude mice.Migration and invasion of renal cell carcinoma cells before and after lncGABPB1-AS1 overexpression were detected by Transwell assays.2.The miRNA with lncGABPB1-AS1 binding site and the target gene downstream of miRNA were predicted by biological website,and the target relationship was verified by double luciferase reporter gene experiment.The miR-1246 expression after lncGABPB1-AS1 overexpressing was detected by RT-PCR;detection of PCK1 expression before and after mir-1246 overexpression by RT-PCR and Western blot;detection of PCK1 expression after miR-1246 and lncGABPB1-AS1 overexpression;CCK8 and Transwell assays were used to detect the proliferation,migration and invasion of renal cell carcinoma cells after overexpression of lncGABPB1-AS1 alone or simultaneous lncGABPB1-AS1 overexpression and PCK1 knockout.The PCK1 expression was detected by immunohistochemistry or Western blot.Results: The lncGABPB1-AS1 expression was negatively correlated with tumor size,TNM stage and Fuhrman grade.After transfection of lncGABPB1-AS,lncGABPB1-AS1 was highly expressed in RCC cells,the ability of proliferation,migration and invasion of RCC cells decreased.Double luciferase reporter gene assays proved that lncGABPB1-AS1 and miR-1246 could bind and miR-1246 and PCK1 could bind.After overexpression of lncGABPB1-AS1,the miR-1246 expression decreased.After overexpression of mir-1246,of PCK1 expression decreased.PCK1 expression in miR-1246 overexpressed group decreased significantly,while it changed little in miR-1246 and lncGABPB1-AS1 overexpressed group.The PCK1 expression was relatively low in renal cancer cells and tissues.After knockdown of PCK1,the proliferation,migration and invasion of renal cancer cells overexpressing lncGABPB1-AS1 increased.Conclusion: 1.The lncGABPB1-AS1 expression in ccRCC and cell lines was significantly lower than the adjacent tissues and normal renal tubular epithelial cells;lncGABPB1-AS1 expression was negatively correlated with TNM stage,tumor size and Fuhrman grade;overexpression of lncGABPB1-AS1 could inhibit the proliferation,migration and invasion of renal carcinoma cells,and inhibit the subcutaneous tumorigenesis of xenotransplantation mice.2.LncGABPB1-AS1 inhibit the proliferation,migration and invasion of renal cancer cells by regulating miR-1246 / PCK1 axis.