Figure 7

miR-125b-5p mimics antagonize MM tumor growth in vivo. In vivo growth of luciferase gene-marked NCI-H929 xenografts intra-tumorally treated with miR-125b-5p mimics or scr controls. Palpable subcutaneous tumor xenografts were treated with 20 μg of NLE-formulated oligos. Intra-tumor injections were administered every other day, for a total of six injections (indicated by arrows). (a) BLI-based measurement of tumor volumes (three mice for each group) was made at 25 days from treatment. (b) Tumors were also measured with an electronic caliper every other day (five mice for each group). Averaged tumor volume of each group ±s.d. is shown. P-values were obtained using two-tailed t-test. (c) Survival curves (Kaplan–Meier) of intra-tumorally treated mice show prolongation of survival in miR-125b-5p-treated NCI-H929 xenografts compared with controls (log-rank test, P<0.05). Survival was evaluated from the first day of treatment until death or sacrifice. Percentage of mice alive is shown. (d) In vivo tumor growth of NCI-H929 xenografts i.p. treated with NLE-formulated miR-125b-5p or miR-NC. I.p. injections were administered twice weekly, for a total of four injections (indicated by arrows). Tumors were measured with an electronic caliper every other day (five mice for each group). Averaged tumor volume of each group ±s.d. is shown. P-values were obtained using two-tailed t-test. (e) Survival curves (Kaplan–Meier) of i.p.-treated mice show prolongation of survival in miR-125b-5p–treated NCI-H929 xenografts compared with controls (log-rank test, P<0.05). Survival was evaluated from the first day of treatment until death or sacrifice. Percentage of mice alive is shown. (f) qRT-PCR of miR-125b-5p expression in lysates from retrieved NCI-H929 xenografts intra-tumorally treated with miR-125b-5p or miR-NC. The results shown are average miRNA expression levels after normalization with RNU44 and ΔΔCt calculations. Data represent the average ±s.d. of three independent experiments. (g) Western blot analysis of BLIMP-1 and IRF4 in lysates from a representative retrieved NCI-H929 xenograft intra-tumorally treated with miR-125b-5p or miR-NC. GAPDH was used as protein loading control. (h) Western blot analysis of CASP10, cFLIP and c-Myc in lysates from a representative retrieved NCI-H929 xenograft intra-tumorally treated with miR-125b-5p or miR-NC. GAPDH was used as a protein loading control.