THAP9-AS1 Promotes Tumorigenesis and Reduces ROS Generation through the JAK2/STAT3 Signaling Pathway by Increasing SOCS3 Promoter Methylation in Osteosarcoma

Figure 2

THAP9-AS1 promoted SOCS3 promoter methylation by recruiting DNMTs. (a) Results of BLAST comparison between THAP9-AS1 and the SOCS3 promoter region. (b) The expression of THAP9-AS1 was significantly upregulated in MG63 cells after transfection with THAP9-AS1-expressing vectors. (c) The binding of THAP9-AS1 to the SOCS3 promoter was evaluated by dual-luciferase reporter gene assay. (d) Fold enrichment of DNMT1, DNMT3a, and DNMT3b in the SOCS3 promoter was determined by ChIP assay. (e) RIP was used to detect the binding between THAP9-AS1 and DNMTs. (f) MSP was conducted to verify the methylation status of SOCS3. U: unmethylation; M: methylation. (g) The results of the BSP assay validated that overexpression of THAP9-AS1 induced promoter hypermethylation of SOCS3; black circle, methylated CpG sites; white circle, unmethylated CpG sites. THAP9-AS1- and THAP9-AS1-expressing vectors; vector, empty vector control. pSOCS3-mut: mutants of the SOCS3 promoter; pSOCS3-wt: wild-type SOCS3 promoter. ; ; .

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