Fig. 5

FOXM1 regulates KIF20A gene transcription in PCa. a, b FOXM1 expression was knocked down in VCaP-DR and DU145-DR cells using siRNA. qRT-PCR and WB were used to measure KIF20A mRNA and protein levels. c, d VCaP cells were transfected with siFOXM1 or in combination with pc-KIF20A. MTT was used to assess cell proliferation. WB was used to analyze levels of cyclinA2, cyclinD1, cyclinE1, bcl-2, and PARP protein in the absence or presence of DOC (12.5 μM). e FOXM1 expression was upregulated by pc-FOXM1. KIF20A was then detected using WB. f–h FOXM1 knockdown in VCaP-DR cells decreased KIF20A gene promoter activity. The FOXM1 protein-bound DNA in VCaP cells was purified using ChIP. Primers spanning the FKH binding motif of the KIF20A gene promoter were designed. FOXM1 binding to the human KIF20A promoter was examined using ChIP-PCR. Protein quantitative analysis was undertaken using the Gel-Pro 32 software, Data represents the mean ± SD (n = 3). ^*p < 0.05, ^**p < 0.01