CCL21 Facilitates Chemoresistance and Cancer Stem Cell-Like Properties of Colorectal Cancer Cells through AKT/GSK-3β/Snail Signals

Figure 4

Snail is crucial for the CCL21 promoted chemoresistance. (a) HCT116 cells were transfected with control vector pcDNA-3.1 (Vector) and pcDNA-Snail for 24 h, and the expression of Snail at protein level was detected by western blotting. (b) HCT116 cells were transfected with control vector pcDNA-3.1 (Vector) and pcDNA-Snail for 24 h, and the expression of Snail at mRNA level was detected by qRT-PCR. (c) Control vector pcDNA-3.1 (Vector) and pcDNA-Snail were transfected into HCT116 cells for 48 h; the cell sensitivity to DOX was detected by MTT. (d) Control vector pcDNA-3.1 (Vector) and pcDNA-Snail were transfected into HCT116 cells for 48 h, and the cell sensitivity to 5-FU was detected by MTT. (e) Control vector pcDNA-3.1 (Vector) and pcDNA-Snail were transfected into HCT116 cells for 48 h, and the expression of P-gp at mRNA level was analyzed via qRT-PCR. (f) Control vector pcDNA-3.1 (Vector) and pcDNA-Snail were transfected into HCT116 cells for 48 h, and the expression of P-gp at protein level was analyzed via western blotting. (g) Control vector pcDNA-3.1 (Vector) and pcDNA-Snail were transfected into HCT116 cells for 48 h, and then RH123 (5 μM) was added to the cells for 2 h. The cells were washed five times with cold PBS, and pictures were acquired by fluorescence microscopy. (h) Control vector pcDNA-3.1 (Vector) and pcDNA-Snail were transfected into HCT116 cells for 48 h, and then Rh123 (5 μM) was added to the cells and incubated for 2 h. The cells were washed five times with cold PBS and digested, and we used the flow cytometry to detect fluorescent intensity.

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