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Research Paper Volume 12, Issue 2 pp 1237—1255

AR-induced ZEB1-AS1 represents poor prognosis in cholangiocarcinoma and facilitates tumor stemness, proliferation and invasion through mediating miR-133b/HOXB8

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Figure 4. ZEB1-AS1 was induced by transcription factor AR. (A) AR sequence and binding sites (E1, E2 and E3) to ZEB1-AS1 promoter region were predicted by using JASPAR database (http://jaspar.genereg.net/). (B) The pcDNA3.1-AR amplified AR mRNA expression in QBC939 and CCLP-1 cells compared with empty vector. (C) Upregulated AR facilitated ZEB1-AS1 expression in QBC939 and CCLP-1 cells corroborated by qRT-PCR. (D) ChIP assays were performed to confirm the direct binding of AR to ZEB1-AS1 promoter in QBC939 and CCLP-1 cells. (E) Luciferase reporter plasmids were constructed with TFBS E2, including wild type and mutant type. (F) The luciferase activity of TFBS E2 WT was markedly promoted by pcDNA3.1-AR cotransfection compared with controls in QBC939 and CCLP-1 cells. ***P < 0.001.