Long Non-coding RNA BZRAP1-AS1 Silencing Suppresses Tumor Angiogenesis In Hepatocellular Carcinoma By Mediating THBS1 Methylation

Background:Hepatocellular carcinoma(HCC)is the most frequent primary liver cancer with a high mortality.Long non-coding RNAs(LncRNAs)have recently emerged as regulators in the development and progression of cancers,and can serve as new therapeutic targets.In the present study,we aim to investigate the potential effect of LncRNA BZRAP1-AS1 on the angiogenesis of HCC.Methods:Microarray-based data analysis was initially employed to screen the differentially expressed HCC-related LncRNAs and genes.The expression of BZRAP1-AS1 and thrombospondin-1(THBS1)in HCC tissues and cells were then determined using RT-qPCR.The gene methylation level was measured by methylation-specific PCR(MSP)and bisulfite sequencing PCR(BSP)assays.Next,the interactions between BZRAP1-AS1,DNA methyltransferase 3B(DNMT3b),and THBS1 were assessed by RIP,RNA pull-down and ChIP assays.Moreover,the roles of BZRAP1-AS1,DNMT3b and THBS1 in angiogenesis in vitro as well as tumorigenesis in vivo were evaluated by the gain-and loss-of function experiments.Results:BZRAP1-AS1 was determined as a highly expressed LncRNA in HCC tissues and cells.Down-regulation of BZRAP1-AS1 in HCC cells inhibited HUVEC proliferation,migration and angiogenesis.By interacting with DNMT3b,BZRAP1-AS1 induced methylation of the THBS1 promoter and inhibited the transcription of THBS1,resulting in promoted angiogenesis of HUVECs.Moreover,silencing of BZRAP1-AS1 repressed the angiogenesis as well as the tumor growth of HCC in vivo via up-regulating THBS1.Conclusion:This study provides evidence that angiogenesis in HCC is hindered by silencing of BZRAP1-AS1.Thus,BZRAP1-AS1 may be a promising marker for the treatment of HCC.