Fig. 5: Circp53–209aa activates the mitochondrial apoptosis pathway in colorectal, lung, stomach, liver and breast cancer cells. | Experimental & Molecular Medicine

Fig. 5: Circp53–209aa activates the mitochondrial apoptosis pathway in colorectal, lung, stomach, liver and breast cancer cells.

From: Extracellular vesicle-mediated delivery of circp53 suppresses the progression of multiple cancers by activating the CypD/TRAP/HSP90 pathway

Fig. 5: Circp53–209aa activates the mitochondrial apoptosis pathway in colorectal, lung, stomach, liver and breast cancer cells.

a, The RNA levels of circp53 and linear p53 were determined by RT–PCR with and without RNase R treatment in RKO, HCT116, HepG2, SGC-7901, A549 and MCF-7 cells. b, The RNA levels of circp53 and linear p53 were determined by RT–qPCR with and without RNase R treatment in RKO, HCT116, HepG2, SGC-7901, A549 and MCF-7 cells. c, The levels of circp53 were significantly lower in patients with CRC compared with NP, as evaluated by RNAscope analysis. Representative staining images are shown, with positive reactions indicated by red arrows. d, Statistical analysis of RNAscope analysis (P < 0.01) .e, RNA levels of circp53 were determined by RT–PCR. f, WB analysis was performed to examine overexpression of circp53–209aa in RKO, HCT116, HepG2, SGC-7901, A549 and MCF-7 cells using the HA-tag antibody. g, The specific peptides from circp53–209aa were identified by MS analysis. h The MTT assay demonstrated decreased cell proliferation rates of circp53-OE RKO, HCT116 cells compared with Ctrl cells. i, The MTT assay demonstrated decreased cell proliferation rates of circp53-OE HepG2, SGC-7901 cells compared with Ctrl cells. j, The MTT assay demonstrated decreased cell proliferation rates of circp53-OE A549 and MCF-7 cells compared with Ctrl cells. k The EdU incorporation assay demonstrated the numbers of proliferating cells were significantly decreased in circp53–209aa-OE cells compared with Ctrl cells. l, Statistical analysis of EdU incorporation assay. m, The effects of circp53 overexpression on cell apoptosis in RKO, HCT116, HepG2, SGC-7901, A549 and MCF-7 cells. n, Statistical analysis of cell apoptosis. o, The TUNEL assay showed the numbers of apoptotic cells were significantly increased in circp53–209aa-OE cells compared with Ctrl cells. p, Statistical analysis of TUNEL assay. q, Flow cytometry showed an increased level of circp53–209aa, resulting in a decrease in the ATP level in cancer cells. r, Confocal microscopic analysis revealed a rapid and almost complete loss of Calcein fluorescence signal in A549, HepG2, SGC-7901 and MCF-7 circp53–209aa-OE cells. s, Statistical analysis of confocal microscopic analysis in A549, HepG2, SGC-7901 and MCF-7 circp53–209aa-OE cells. t, Confocal microscopic analysis revealed a rapid and almost complete loss of Calcein fluorescence signal in RKO and HCT116 circp53–209aa-OE cells. u, Statistical analysis of confocal microscopic analysis in RKO and HCT116 circp53–209aa-OE cells. v, The effects of circp53 overexpression on the mitochondrial apoptotic pathway associated proteins Bad, Bak, Bcl-xL, Bax and Bcl-2 in RKO and HCT116 cells. w, The effects of circp53 overexpression on the mitochondrial apoptotic pathway associated proteins Bcl-xL and Bcl-2 in HepG2 and SGC-7901 cells. x, Statistical analysis of expression of mitochondrial apoptotic pathway associated proteins Bad, Bak, Bcl-xL, Bax and Bcl-2 in RKO and HCT116 cells. y, Statistical analysis of expression of mitochondrial apoptotic pathway associated proteins Bad, Bak, Bcl-xL, Bax and Bcl-2 in HepG2 and SGC-7901 cells. The data are presented as mean ± s.d. (n = 6 clinical samples for each group, n = 3 cultures for each group, *P < 0.05, **P < 0.01 and ***P < 0.001).

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