Fig 1: p53 mutation enhances the resistance of CRC cells to PDT. p53 wt RKO and p53 mut SW480 cells were treated or non-treated with PDT and examined for cell viability by CCK-8 assay (A), cell apoptosis by Flow cytometry (B), and the expression levels of c-Myc (C), NEAT1 (D), miR-124 (E), and iASPP (F) by qRT-PCR. *P < 0.05, **P < 0.01, compared to RKO cells. ## P < 0.01 compared to RKO+PDT group.
Fig 2: Schematic diagram of the role of p53 mutation or knockout in PDT treatment. In the present study, we revealed that p53 promoted miR-124 expression to inhibit iASPP expression, so as to amplify the inhibitory effect of PDT on CRC cell proliferation; after p53 mutation or knockout, miR-124 expression was downregulated while the iASPP expression was upregulated, so that the inhibitory effect of PDT on CRC cell proliferation was reduced
Fig 3: A schematic mechanism. In p53wt CRC cells, PDT inhibits the c-Myc/NEAT1 axis and promotes the miR-124/iASPP/p53 feedback loop that amplifies PDT functions, inducing cancer cell apoptosis. In p53mut CRC cells, the expression of the c-Myc/NEAT1 axis was up-regulated, and the miR-124/iASPP/p53 feedback loop could not be activated, leading to the failure in inducing apoptosis and, thus, PDT tolerance.
Fig 4: The correlation of p53, miR-124 and iASPP expression in CRC tissues. (a) Clinic samples containing diverse p53 levels have been obtained and the pathological states have been assessed. Results showed that the samples possessing higher p53 content had a better pathological condition. (b) miR-124 expression was upregulated in p53wt cells but downregulated in p53mut cells, and the expression levels were altered in a p53 content-dependent manner; iASPP expression was downregulated in p53wt cells but upregulated in p53mut cells, and the expression levels were altered in a p53 content-dependent manner. The data are presented as mean±S.D. of three independent experiments. **P<0.01, ***P<0.005
Fig 5: The suppressing effect of miR-150 on CRC cells depends on the overexpression of iASPP. Overexpression of iASPP increases cell viability and inhibits cell cycle arrest and apoptosis in SW480 and HCT116 cells. (A) iASPP was decreased after the cells were transfected with mimics. (B) CCK-8 assay exhibited that downregulation of iASPP increased the OD450 value at 48 and 72 h when compared with the mimics group. (C) Flow cytometry assay demonstrated that a lower number of cells distributed in the G1 phase was recorded in the mimics+pcDNA group when compared with the miRNA group. (D) Hoechst staining indicated that less Hoechst-positive cells were observed in the mimics+pcDNA group. Blank, SW480 cells; NC, negative control; Mimics, miR-150 mimics. **P<0.05 vs. mimics.
Supplier Page from Abcam for Anti-iASPP antibody