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

Fig 7. High CCL20 predicted poor prognosis of breast cancer patients, and the blockade to CCL20-NF-κB pathway reversed the chemoresistance of breast cancer.

Fig 7

(A-B) The OS (A) and RFS (B) influenced by CCL20 expression in breast cancer from the online database (PROGgeneV2). Grades 1–3, the histological grading of breast cancer. (C-D) Nude mice were injected with 1 × 106 shCTRL or CCL20-knockdown (sh2, sh5) MDA-MB-231 cells at fourth mammary fat pads. When the average tumor volume reached 20 mm3, mice of each group were divided into 2 new groups randomly at similar tumor volume and treated with DOC (10 mg/kg, i.p., once a week) or vehicle (“veh.”). Tumor size was monitored once a week, (C) and tumor image was taken after mice were killed (D). Tumor number n = 4. (E) Tumors from (D) were minced and digested into single cells for determining the ALDH+ BCSC population by flow cytometry. (F-G) Nude mice were injected with 1 × 105 cells digested from PDX of TNBC (#USTC11 established by our group) at fourth mammary fat pads. After tumor initiation, mice were divided into 4 groups as criteria in (C) and treated with vehicle, DOC (10 mg/kg), CAPE (10 mg/kg), or both once a week i.p. Tumor size was monitored every 3 days (F), and tumor image was taken after mice were killed (G). Tumor number n = 4. (H) ALDH+ population in tumors from (G) was determined as method in (E). Four tumors per group (except CAPE group, in which 1 tumor was too small to obtain enough single cells for determination). These experiments were repeated at least twice. Data are shown as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001 analyzed by unpaired t test (E and H) or multiple comparison test of 2-way ANOVA (C and F) and log rank test (A and B). ALDH, aldehyde dehydrogenase; BCSC, breast cancer stem cell; CAPE, caffeic acid phenethyl ester; CCL20, C-C motif chemokine ligand 20; DOC, docetaxel; i.p., intraperitoneally; OS, overall survival; PDX, patient-derived xenograft; RFS, relapse-free survival; shCTRL, scramble control; TNBC, triple-negative breast cancer.