Skip to main content
. Author manuscript; available in PMC: 2022 Aug 9.
Published in final edited form as: Oncogene. 2021 Nov 20;41(5):671–682. doi: 10.1038/s41388-021-02118-4

Figure 5. CSN5i-3 inhibited oncogenic activity of CSN5 in prostate cancer cells.

Figure 5.

A, CSN5i-3 efficacy in 7 prostate cancer cells. Cell proliferation assay was performed after 3 days of treatment with CSN5i-3. B, GSEA of cell cycle and E2F_targets pathway in CSN5i-3 treated C4–2 cells. C and D, The regulation of AR signaling by CSN5i-3. AR signaling enrichment was analyzed by GSEA in C4–2 (1μM CSN5i-3) and LNCaP (5μM CSN5i-3) cells with CSN5i-3 treatment for 3 days (C). AR protein and AR targets were detected by western blot in C4–2, LNCaP, and 22Rv1 (5μM CSN5i-3) cells with either CSN5i-3 or MLN4924 (1μM) for 3 days (D). E and F, DNA damage evaluation with CSN5i-3 treatment. DNA repair pathway enrichment in CSN5i-3 treated C4–2 (E). DNA damage marker activation was monitored in C4–2 cells using Muse multicolor DNA damage kit (F). G, Apoptosis induced by CSN5i-3 treatment. Apoptosis was detected by Annexin V assays and immunoblots in C4–2 (1μM CSN5i-3) and PC3 (10μM CSN5i-3) cells. H, p53 signaling and apoptosis pathways enriched in C4–2 cells with CSN5i-3 treatment. I, Four oncogenic pathways were enriched in CSN5i-3 downregulated gene group in LNCaP, C4–2, 22Rv1, and PC3 cells. J, Invasion assays with CSN5i-3 treated PC3 cells. Invasion ability was detected after cells were treated with CSN5i-3 for 72 hours. K, EMT_up and metastasis_up pathway enrichment with CSN5i-3 treatment in PC3 cells. L, Regulation of prostate cancer SRGs by CSN5i-3 in PC3 cells: 36 genes among 186 prostate cancer SRGs were inhibited by CSN5i-3 in PC3; 4 genes of 13 unfavorable SRGs were detected by qRT-PCR in CSN5i-3 treated PC3 cells. M, Phosphokinase array analysis after 48-hour CSN5i-3 treatment in C4–2 cells. The whole-cell lysates were collected for human phosphokinase array analysis. Relative phosphorylation of spots was quantified by Image J software, and the value of vehicle (NC) was set up as “1.” Figure values represent the mean ± SE of three independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001; vs. control groups (NC) treated with DMSO.