Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Chemotherapy-elicited exosomal miR-378a-3p and miR-378d promote breast cancer stemness and chemoresistance via the activation of EZH2/STAT3 signaling

doi: 10.1186/s13046-021-01901-1

Figure Lengend Snippet: The expression of miR-378a-3p and miR-378d in serum exosomes is increased in breast cancer patients receiving chemotherapy and associated with chemoresistance. a Scanning electron microscopy of serum-exosomes isolated from patient serum. b Western blot analysis of the key enriched proteins CD81 and TSG101 in breast cancer patient serum exosomes. c Sequencing analysis of exosomal microRNAs from patient serum exosomes before receiving neoadjuvant chemotherapy, after receiving one cycle of neoadjuvant chemotherapy and after receiving four cycles of neoadjuvant chemotherapy. d Exosomes were isolated from the serum of breast cancer patients before and after neoadjuvant chemotherapy ( n = 24). Variation in serum exosomal miR-378a-3p and miR-378d in groups with different responses to neoadjuvant chemotherapy. e Five-year survival rates in patients with low or high expression miR-378 based on the TCGA database ( n = 854). f Analysis of miR-378 expression and the corresponding novel stemness index mRNAsi in 673 breast cancer patients in the TCGA database. g Pre- and post-chemotherapy expression data were analyzed by Gene set variation analysis for the breast cancer cohort ( n = 10). h Western blot analysis of protein expression in breast cancer tissue before and after neoadjuvant chemotherapy in the no response group ( n = 12). i The expression level of miR-378 was positively correlated with β-catenin , NOTCH1 or EZH2 in breast cancer ( n = 470). * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: The data after subsequent miRNA RT-qPCR analysis (Qiagen, USA) were standardized at the level of this spiked control.

Techniques: Expressing, Electron Microscopy, Isolation, Western Blot, Sequencing

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Chemotherapy-elicited exosomal miR-378a-3p and miR-378d promote breast cancer stemness and chemoresistance via the activation of EZH2/STAT3 signaling

doi: 10.1186/s13046-021-01901-1

Figure Lengend Snippet: Chemo-elicited exosomal miR-378a-3p and miR-378d regulated the WNT/β-catenin and Notch stemness pathways by targeting NUMB and DKK3. a Analysis of miRNA-378 expression data by Gene set enrichment analysis revealed that genes involved in the WNT and NOTCH pathways were significantly enriched. b MiRanda prediction of the binding sites of miR-378a-3p/miR-378d within the 3′-UTR of DKK3/NUMB . c , d Relative levels of DKK3/NUMB mRNA in CAL51 cells transfected with miR-378a-3p or miR-378d mimics and the corresponding normal control. e–h MiR-378a-3p/miR-378d was found to be directly bound to the NUMB/DKK3 3′-UTR using a dual-luciferase reporter assay. i, j Western blot analysis of the expression of DKK3, β-catenin, Cyclin D1, C-MYC, NUMB, NOTCH1, HES1 and HEY1 in the different groups. i CAL51 and MDA231 cells were transfected with miR-378a-3p or miR-378d mimics, inhibitors and the corresponding normal control for 48 h before western blot analysis. j Western blot results showing the expression levels of the indicated proteins in CAL51 and MDA231 cells at 48 h after simultaneous transfection with the DKK3 or NUMB expression plasmid or empty vector and miR-378a-3p or miR-378d mimics or the corresponding normal control. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: The data after subsequent miRNA RT-qPCR analysis (Qiagen, USA) were standardized at the level of this spiked control.

Techniques: Expressing, Binding Assay, Transfection, Control, Luciferase, Reporter Assay, Western Blot, Plasmid Preparation