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

Figure 3

Snail is vital to chemoresistance and cancer stem cell properties promoted by CCL21. (a) HCT116 cells were treated with CCL21 (100 ng/mL) for 2, 4, 8, 12, 24, 48, and 72 h and the expression of Snail at protein level was analyzed by western blotting. (b) HCT116 cells were treated with CCL21 (100 ng/mL) for 24 h, and the expression of Snail at mRNA level was analysed by qRT-PCR. (c) SiNC or si-Snail siRNAs were transfected into cells for 24 h and treated with or without CCL21 (100 ng/mL) for 24 h, and then the expressions of Snail were detected by western blotting. (d) SiNC or si-Snail siRNAs were transfected into cells for 24 h and treated with or without CCL21 (100 ng/mL) for 72 h, and then the cell sensitivity to DOX was detected by MTT. (e) SiNC or si-Snail siRNAs were transfected into cells for 24 h and treated with or without CCL21 (100 ng/mL) for 72 h, and then the cell sensitivity to 5-FU was detected by MTT. (f) SiNC or si-Snail siRNAs were transfected into cells for 24 h and treated with or without CCL21 (100 ng/mL) for 72 h, and then RH123 (5 μM) was added to cells for 2 h. Following that, the fluorescent intensity was analyzed via flow cytometry. (g) SiNC or si-Snail siRNAs were transfected into cells for 24 h and treated with or without CCL21 (100 ng/mL) for 72 h, and then the expressions of P-gp, Bmi-1, Nanog, and OCT-4 were detected by western blotting. (h) SiNC or si-Snail siRNAs were transfected into cells for 24 h and treated with or without CCL21 (100 ng/mL) for a week, detecting the mammosphere forming ability.

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