Figure 3: Systemic delivery of antagomiR-135b inhibits metastasis and tumour growth in vivo.

(a) Nude mice were subcutaneously injected with 5 × 10⁵ H441 cells. Scramble antagomir (NC) or miR-135b Antagomir (antago-135b) was administered via tail vein injection 2 days after implantation. On day 14 after cancer cell implantation, tumour volume measurements began and were performed every 4 days (n=9). Mean±s.e.m. is shown, *P<0.05 by Student’s t-test; (b–d) CL1-5 cells were implanted orthotopically into the left lungs of 6-week-old nude mice (n=9). A series of scramble antagomir (NC) or miR-135b antagomir (Antago-135b) intravenous injections were performed every 2 days after tumour cell implantation. Lungs were harvested 25 days after the cells were implanted. (b) Bright-field imaging and H&E staining of CL1-5 tumour-bearing lungs. Arrows indicate the visible nodules. (c) Primary tumour volume and (d) number of intra-lung metastatic nodules in CL1-5 tumour-bearing mice treated with scramble antagomir (NC) or miR-135b antagomir (Antago-135b) 25 days after orthotopic implantation. Data are expressed as the mean±s.e.m. *P<0.05 by Student’s t-test. (e) Mice bearing CL1-5 cells were subjected to scramble antagomir (NC) or miR-135b antagomir (Antago-135b) treatment. Animal survival was determined using Kaplan–Meier analysis and the log-rank test for data obtained 25 days after the cancer cells were orthotopically injected. (n=9) (f) CL1-5-F4 cells with scramble antagomir (NC) or miR-135b antagomir (Antago-135b) were intravenously injected to assess the effect of antagomir on late-stage metastasis. (n=9). Representative average of visible metastatic lung nodules and H&E staining (right). Data are expressed as the mean±s.e.m. *P<0.05 by Student’s t-test.