Figure 2.

Antiandrogen Enz treatment suppresses BRCA1 and HR pathway. A) The protein expression correlation between AR and BRCA2 and RAD51 in 184 HCC patients based on TCGA database. B) HA22T cells were infected with lentivirus pLKO control or shRNA targeting AR and western blots showed some key HR proteins expression levels (left). SK-HEP-1 cells were infected with lentiviruses pWPI control or oeAR and western blots showed proteins expression levels of BRCA1 and BCL2. BCL2 protein level was quantified using ImageJ (right upper). HA22T cells were treated with 10 μM Enz (ENZ), western blots showed proteins expression levels of BRCA1 (right lower). C) HA22T cells were treated with DMSO or 20 μM ENZ for 24 h, then irradiated by 4 Gy IR. Three hours after radiation, DNA damage was detected by Alkaline CometAssay^®. The comet tails were scored according to DNA content. Multiple randomly selected cells were analyzed per sample. Scale bar 200 μm. D) HA22T cells were pretreated with DMSO or 20 μM ENZ for 24 h, followed by 4 Gy IR. 24 h after radiation, r-H2AX and GAPDH were detected by western blot. E) SK-HEP-1 cells were pretreated with DMSO or 20 μM Enz for 24 h and were co-transfected with pDRGFP and I-SceI plasmids. After 48 h, the GFP+ cells were observed under fluorescence microscope. Scale bar 100 μm. The proportion of GFP+ cells were presented as relative percentage. Five randomly selected microscopic fields were analyzed per sample. For B, C, and E, quantitation is at the far right. Data are presented as mean ± SD. **P < 0.01, NS=Not Significant.