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. 2020 Jul 7;13(7):144. doi: 10.3390/ph13070144

Figure 1.

Figure 1

miR-101-3p levels are downregulated in brain metastatic breast cancer cells and vary inversely with their brain metastatic ability. (A) The expression profile of miR-101-3p was examined by real-time PCR in three breast cancer cell lines with different brain metastatic propensities (MCF-7, MDA-MB-231-TGL and MDA-MB-BrM2). Relative miR-101-3p level expression was normalized against the U6 small nuclear RNA levels. (B) The transmigration abilities of the different BC cells were examined by trans-endothelial migration assay. The amount of transmigrated cells was determined by fluorescence measurements.(C) The relative mRNA expression profile of three pro-metastasis genes known to mediate brain trans-endothelial migration of BC cells (PTGS2 coding for COX-2, ST6GALNAC5 and HBEGF) was examined in the three BC cell lines using real time PCR and normalized against GAPDH mRNA levels. (D,E) Western Blot analysis (D) and immunofluorescence images (E) of COX-2, ST6GALNAC5 and HBEGF in MCF-7, MDA-MB-231-TGL and MDA-MB-BrM2 cell lines. Experiments were carried out three times. Data are mean ±SD from three independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001.