Figure 1.
TNTα-miR-126 inhibits murine HE tumor growth in vivo
(A) Schematic diagram of in vivo tissue nanotransfection technology-mediated delivery of oligonucleotides based on the voltage gradient. (B) RNA in situ detection of miR-126 in the HE tumor sections (d10) treated with scrambled oligonucleotides (TNTsham) or antagomir-126 (TNTα-miR-126). (C) Intensity calculation of RNA in situ images determined the significant reduction of miR-126 level in the TNTα-miR-126 group compared with TNTsham. ∗p < 0.05 (n = 4), Mann Whitney test. (D) Level of miR-126 from the biopsy samples isolated from non-tumor (NT), HE tumor (T), and skin from HE tumor region treated with antagomir-126 (T + TNTα-126). ∗p < 0.05 (n = 4), Mann Whitney test. (E) Representative images of 129P3/J mice treated with TNTsham and TNTα-miR-126 at d12 and d29 showing decrease in tumor growth in TNTα-miR-126-treated mice. (F) Tumor volume of day-matched animals for the treatment of TNTsham and TNTα-miR-126. ∗p < 0.05 (n = 5), Mann Whitney test. (G) TNT-based delivery (twice a week for 4 weeks) of antagomir-126 (red) inhibits HE progression. Tumor volume was quantified using calipers (length ×·width ×·height). In TNTsham mice, tumor volume sharply rose, causing death on d12–d17. (H) Kaplan-Meier survival curve of TNTsham and TNTα-miR-126 groups monitored for 90 days shows tumor-free survival for TNTα-miR-126 group. (I) Ultrasound imaging of animals from each group (n = 5) was performed on a Vevo-2100 system using high-frequency linear array transducers operating between 8 and 17 MHz. (J) Tumor blood flow analyzed using Vevo-2100 system shows significant difference at d10 after TNTα-miR-126 treatment compared with TNTsham groups. (K) H&E images show complete regression of tumor with TNTα-miR-126 treatment on d29 compared with d12 TNTα-miR-126 group. (L) Day-matched immunohistochemical analysis showing decrease in ratio of CD31+ (red) and LYVE1+ (green) cells in TNTα-miR-126-treated HE tumor at d12 compared with TNTsham group-treated HE tumor, and (M) its quantification. ∗p < 0.05, n = 5, Mann Whitney test. Data presented as mean ± SEM.