Fig 1: CCL20/CCR6 enhances the stemness and tumorigenicity of CD44+CD117+ cells treated with PTX via the Notch1 signaling pathway. (A) Expression level of Ki-67 in tumor was determined via immunohistochemistry. (B) Apoptosis of tumor cells was determined using a TUNEL assay. (C) Reactive oxygen species levels of tumors were determined by the dichlorofluorescein diacetate method. Scale bar, 100 µm. (D) Expression levels of Oct4, CCR6, Notch1 and ABCG1 in tumor tissue, as determined via western blotting. (E) Semi-quantitative analysis of Oct4, CCR6, Notch1 and ABCG1 protein expression. ****P<0.0001 vs. CD44+CD117+ group; ####P<0.0001 vs. CD44−CD117− group; $$$$P<0.0001 vs. CD44+CD117+ + CCL20 group. PTX, paclitaxel; CCL20, C-C motif chemokine ligand 20; CCR6, CCR6, C-C motif chemokine receptor 6; Notch1, notch receptor 1; ABCG1, ATP binding cassette subfamily G member 1.
Fig 2: CD44+CD117+ cells have a powerful PTX resistance compared with CD44-CD117- cells. (A) Expression levels of Oct4 and CCR6 were determined via reverse transcription-quantitative PCR. (B) Sphere formation analysis of CD44+CD117+ and CD44-CD117- cells. (C) Expression levels of Oct4, CCR6, Notch1 and ABCG1 were determined via western blotting. (D) Semi-quantitative results of protein expression. (E) IC50 of CD44+CD117+ and CD44-CD117- cells to PTX at 24 and 48 h. ***P<0.001 and ****P<0.0001 vs. CD44-CD117- group. PTX, paclitaxel; CCR6, CCR6, C-C motif chemokine receptor 6; Notch1, notch receptor 1; ABCG1, ATP binding cassette subfamily G member 1.
Fig 3: siNotch1 reverses the effect of the CCL20/CCR6 axis in enhancing the stemness of CD44+CD117+ cells. (A) Expression level of Oct4, as determined by reverse transcription-quantitative PCR. (B) Protein expression levels of Oct4, CCR6, Notch1 and ABCG1 were determined via western blotting. (C) Semi-quantitative analysis of Oct4, CCR6, Notch1 and ABCG1 protein expression. (D) Sphere formation analysis of CD44+CD117+ cells. Scale bar, 100 µm. ****P<0.0001 vs. Control group; ###P<0.001 and ####P<0.0001 vs. CCL20 group. NC, negative control; si, small interfering RNA; CCR6, CCR6, C-C motif chemokine receptor 6; CCL20, C-C motif chemokine ligand 20; ABCG1, ATP binding cassette subfamily G member 1; Notch1, notch receptor 1.
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