Skip to main content
. 2018 Jul 27;16(7):e2005869. doi: 10.1371/journal.pbio.2005869

Fig 3. CCL20 promoted tumor progression and enhanced the chemoresistance to taxanes in TNBC.

Fig 3

(A) CCL20 level in breast cancer tissues based on the RNA-seq data. Result here was in whole or part based upon data generated by the TCGA Research Network: http://cancergenome.nih.gov/. (B-C) CCL20 levels determined by qRT-PCR or western blot in 7 breast cancer cell lines. (D) MTT assay conducted with MDA-MB-231 cells. shCCL20-2 and shCCL20-5: two CCL20-shRNA. (E) Matrigel invasion assay conducted with MDA-MB-231. Scale bars: 400 μm. (F) Quantitative analysis of total invaded cells in (E). (G) Soft agar colony formation assay performed with MDA-MB-231 cells, and colony numbers were quantified. (H-K) MTT assay, Matrigel invasion assay, and soft agar colony formation assay were carried out with vector control or CCL20-overexpressing MDA-MB-231 cells with the same methods as described in (D-G). Scale bars: 200 μm. (L) MTT assay conducted with MDA-MB-231 cells in the presence or absence of rhCCL20 (10 ng/ml) or anti-CCL20 (200 ng/ml). (M) Matrigel invasion assay conducted in 3 groups as in (L) and quantitative analysis of total invaded cells. (N-O) Nude mice were injected with 1 × 106 vector or CCL20-overexpressing MDA-MB-231 at fourth mammary fat pads. Tumor size was monitored once a week (N), and tumor image was shown after mice were killed (O). Five tumors per group. The experiments were repeated at least twice. (P-Q) Vector control or CCL20-overexpressing MDA-MB-231 (P) and SUM159 (Q) were treated with DOC (MDA-MB-231, 14.10 nM; SUM159, 10 nM) for indicated days, and the chemoresistance was analyzed as described in evaluation of chemoresistance index. Data were shown as mean ± SEM and are representative of 3 individual experiments. *p < 0.05, **p < 0.01, ***p < 0.001 by unpaired t test of triplicates and multiple comparisons test of 2-way ANOVA (D, H, L, and N). anti-CCL20, CCL20 neutralization antibody; CCL20, C-C motif chemokine ligand 20; DOC, docetaxel; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide; qRT-PCR, quantitative real-time PCR; rhCCL20, recombinant human CCL20; RNA-seq, RNA sequencing; shCTRL, scramble control; shRNA, short hairpin RNA; TCGA, The Cancer Genome Atlas; TNBC, triple-negative breast cancer.