Figure 6.
In vivo effect of miR-122 on primary tumour growth and metastasis. (a) Tumour growth curve in mice carrying MCFDCIS/vec and MCFDCIS/miR-122 orthotopic xenografts (n = 7 mice). (b) BLI imaging at week 3. (c) Luciferase quantification of (b) (n = 7 mice). (d) IHC for Ki67, PKM1/2, and GLUT1 in tumour, brain, and lung sections. Scale bar is 100 μm. (e) Quantification of Ki67+ tumour cells from 3 fields per tumour (n = 6 mice per group). (f) Intratumoural levels of ATP were assessed in tumour lysates by ENLIGHTEN ATP assay (n = 6 mice per group). (g) miR-122 levels in the brain and lungs determined by RT-qPCR (n = 18 extracts from 6 mice). (h) 2-NBDG uptake quantification in the tumour, brain, and lungs (n = 12 fields from 4 mice per group). (i–j) Luciferase qPCR in the brain (i, n = 24 extracts from 8 mice per group) and lungs (j, n = 18 extracts from 6 mice per group) of MCFDCIS tumour-bearing mice. (k) Tumour growth curve in mice carrying orthotopic xenografts of MDA-MB-231 with stable knockdown of miR-122 (MDA-MB-231/122KD) and also receiving EV treatments as indicated (n = 7 mice per group). No significant difference (p > 0.05) between groups based on Kruskal-Wallis test. (l–m) Luciferase qPCR for the detection of metastases in the brain (l) and lungs (m) of mice bearing MDA-MB-231/122KD tumours and treated with indicated EVs (n = 12 extracts from 4 mice per group). * p < 0.05, ** p < 0.01, *** p < 0.001 for all panels derived from Kruskal-Wallis test. Data are represented as mean ± SD in all panels except (b & d).