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Fig. 6 | Breast Cancer Research

Fig. 6

From: Mebendazole prevents distant organ metastases in part by decreasing ITGβ4 expression and cancer stemness

Fig. 6

Mebendazole treatment reduces ITGβ4 expression. A Venn diagram displaying the number of genes with differential expression following MBZ treatment (− 2 ≥ FC ≥ 2) that overlap or are exclusive to MDA-MB-231 (pink circle) and SUM159 (blue circle) cells. Additional file 3 includes the gene lists used to generate the VENN Diagram. B A table representing DEGs that are downregulated in both MDA-MB-231 and SUM159 cells and that have been implicated in cancer metastasis is presented with individual log2 fold change from the RNA sequencing analysis C, D Quantification of ITGβ4 relative gene expression from RT-qPCRs performed with SUM159 C and MDA-MB-231 D cells treated for 72 h with indicated doses of MBZ or DMSO. Mean ± SEM; N = 3 independent experiments with n = 3 technical replicates. P values for one-way ANOVA test ****< 0.0001. E Representative histograms for ITGβ4 expression in MDA-MB-231 and SUM159 cells as measured by flow cytometry after 72 h of treatment with 0.5 μM and 1 μM MBZ or DMSO. The APC peak median values are listed. F Expression of ITGβ4 protein analyzed by Western blot using lysate extracted from MDA-MB-231 and SUM159 cell lines treated with increasing doses of MBZ as indicated or DMSO. G Representative FACS analysis of the ITGβ4 + MDA-MB-231 and SUM159 cells that were co-stained with CD44 and CD24 antibodies after being treated with 1 μM of MBZ or DMSO for 72 h. H The fold change of CD44hiCD24lowcells within the ITGβ4+ gated population of SUM159 and MDA-MB-231 cells described in (G). Mean ± SEM; N = 3 independent experiment. P values for paired two-way t test **< 0.01 I, J Quantification of immunofluorescent staining for ITGβ4 protein in tissue sections from mouse tumors (Fig. 4) treated with either 30 mg/kg MBZ in sesame oil suspension (N = 6) via gavage or vehicle control (sesame oil alone; N = 6) with the primary tumor onboard (I) and tumor removal (J). Mean ± SEM; N = 6 or 5 independent mice per group with n = 5 technical replicates for percent positive area quantification. P values for two-way t test * < 0.05

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