Fig 1: Genic or chemical downregulation of Pin1 suppressed cell proliferation in CCCs. (A) Chemical synthesis steps of KPT-6566. (B) Mass spectrum of KPT-6566, ESI-MS: m/z 466.0 [M+Na]+. (C) Hydrogen spectroscopy to confirm chemical structure of KPT-6566, 1H NMR (300 MHz,DMSO-d6) δ 8.14 - 8.04 (m, 2H), 8.03 -7.97 (m, 2H), 7.90 - 7.87 (s, 1H), 7.87 - 7.80 (m, 2H), 7.76 - 7.69 (m, 2H), 3.99 (s, 2H), 1.35 (s, 9H). (D) Cell viability assay of the Hela-shPin1/Hela-shNC and SiHa-shPin1/SiHa-shNC for 24 h. (E) Hela, SiHa or HUVEC cells were treated with KPT-6566 and the growth curves were plotted over concentration. (F) Representative micrographs of the colonies of Hela-shPin1/SiHa-shPin1 were counted and compared with that of NC. (G) Representative micrographs of the colonies of Hela/SiHa treated with KPT-6566 were counted and compared with that of treated with DMSO. Each assay was performed in triplicate. *P<0.05.
Fig 2: Pin1 and TAZ are highly expressed in TNBC. (A) Pin1 and TAZ are highly expressed in breast cancer. (B) Pin1 and TAZ are highly expressed in high-stage breast cancer. (C) Pin1 (N-vs-Luminal: 1.6 × 10-12, N-vs-HER2Pos: 0.18, N-vs-TNBC-BL1: 0.32, N-vs-TNBC-BL2: 0.0093, N-vs-TNBC-IM:0.0079, N-vs-TNBC-LAR: 0.85, N-vs-TNBC-MSL: 0.18, N-vs-TNBC-M:0.024, and N-vs-TNBC-UNS: 0.013.) and TAZ (N-vs -Luminal: 0.00012, N-vs-Her2Pos: 0.57, N-vs-TNBC-BL1: 0.007, N-vs-TNBC-BL2: 0.08, N-vs-TNBC-IM: 0.00068, N-vs-TNBC-LAR: 0.13, N-vs-TNBC-MSL: 0.08, N-vs-TNBC-M: 0.49, and N-vs-TNBC-UNS: 0.093) are highly expressed in TNBC. (D) Relationship between Pin1 and TAZ expression and patient survival in TNBC patients.
Fig 3: KPT-6566 inhibited CC tumor growth by targeting Pin1 in nude mice. (A) Xenograft tumors were harvested 8 weeks after implantation. (B) SiHa tumor volumes were measured weekly for 8 weeks and the curves of tumor volumes were plotted over time. *P < 0.05. (C) Data points are presented as the means ± SD for tumor weights of the tumors were analyzed. *P < 0.05. (D) Expression of Pin1 in xenograft tumors of nude mice were detected by IHC (original magnification × 200). (E) The xenograft tumor sections were subjected to H&E staining, and the percentage of the necrosis areas were counted. *P < 0.05. (F) The percentage of CC cell apoptosis internal xenograft tumor was counted through TUNEL. *P < 0.05. (G) The heart, liver, spleen, kidney and lung sections of combinational treatment nude mice were subjected to H&E staining.
Fig 4: KPT-6566 blocked multiple cancer-driving pathways simultaneously in CCCs. (A-D) Hela/SiHa cells were treated with 5 µM KPT-6566. Expression of Pin1, c-Jun, ß-catenin, cyclinD1, A KT-p473, ERK1/2, p65/NF-?b, GSTP1 and H2AX were detected by western blot assay with specific antibodies. Each assay was performed in triplicate. *P<0.05.
Fig 5: CI regulates the Pin1–TAZ signaling pathway in TNBC cells. (A) Levels of expression of Pin1 and TAZ in MDA-MB-231 cells treated with CI. (B) Levels of expression of Pin1 and TAZ in HCC 1937 cells treated with CI. (C) Levels of expression of Pin1 and TAZ in 4T1 cells treated with CI.
Supplier Page from Abcam for Anti-Pin1 antibody