Figure 5
From: RNA-seq transcriptome analysis of breast cancer cell lines under shikonin treatment
Effect of shikonin on the expression level of DUSP1 and DUSP2 and the activation of MAPKs pathway in breast cancer cells. (A) Different breast cancer cells, MCF-7, SK-BR-3 and MDA-MB231, and human mammary epithelial cells, M10, were incubated with or without shikonin 10 μM for 6 h. The expressions of DUSP1 and DUSP2 were determined by qRT-PCR. Data are presented as mean ± SD from three independent experiments. The statistical significance of the difference between two experimental measurements was assessed by Student’s t-test and represented as follows: ***P < 0.001 vs. shikonin 0 μM (DMSO control). (B) MB-231 cells were treated with different concentrations of shikonin for 6 hr. The expressions of DUSP1 and DUSP2 were analyzed by Western blot. The expression of β-actin was used as a loading control. (C) MB-231 cells were treated with different concentrations of shikonin for 24 hr. The expressions of phospho-JNK 1/2, JNK 1/2, phospho-p38, p38, phospho-ERK1/2, and ERK 1/2 were analyzed by Western blot. The Western blotting results are representative of results obtained in three separate experiments. (D) Gene expression levels of DUSP1 and DUSP2 from TCGA RNA-seq data in many types of cancer were analyzed by DriverDB23,24. The list of abbreviations is shown as follows: BLCA: bladder urothelial carcinoma; BRCA: breast invasive carcinoma; BRCA-T: BRCA-associated triple-negative breast cancer; CESC: cervical squamous cell carcinoma and endocervical adenocarcinoma; CHOL: COAD: colon adenocarcinoma; GBM: glioblastoma multiforme; HNSC: head-neck squamous cell carcinoma; KIRC: kidney renal clear cell carcinoma; KIRP: kidney renal papillary cell carcinoma; LIHC: liver hepatocellular carcinoma; LUAD: lung adenocarcinoma; LUSC: lung squamous cell carcinoma; PAAD: pancreatic adenocarcinoma; PCPG: pheochromocytoma and paraganglioma; PRAD: prostate adenocarcinoma; READ: rectum adenocarcinoma; SARC: sarcoma; SKCM: skin cutaneous melanoma; THCA: thyroid cancer; THYM: thymoma; UCEC: uterine corpus endometrial carcinoma. (E) Shikonin inhibits cell growth and induces apoptosis in different types of breast cancer cells through enhances the expression of DUSP1 and DUSP2 and reduces the activity of their downstream signaling molecules, JNK and p38.
