Figure 2

LncRNA-TUG1 knockdown inhibits tumor growth and angiogenesis in vivo. An in vivo hepatoblastoma model was established by injection of stable TGU1 knockdown HuH-6 cells and scrambled shRNA-transfected HuH-6 cells. (a) TUG1 levels in tumors isolated from the TUG1 knockdown (KD) group and negative control group (Nc) were determined by qRT-PCRs (Student’s t-test, P=0.0011). (b) Comparison of tumor growth between TUG1 KD group and Nc group. The volume of solid tumor was measured every week using a vernier caliper. Tumor volume (V) was calculated using the formula, V= (L × W²) × 0.5 (n=6; Mann–Whitney U-test; P=0.0362 (2w), P=0.0252 (3w), P=0.0182 (4w)). (c) The tumors were weighed immediately after isolation from mice. Tumor weight was plotted between the two groups (Mann–Whitney U-test; P=0.0089). (d and e) CD31 and Ki67 immunofluoresence staining was conducted to detect microvascular density and cell proliferation in hepatoblastoma tissues from TUG1 KD group and Nc group. A representative image and statistical result was shown (n=6; Mann–Whitney U-test; P=0.0045 (d) and P=0.0034 (e)). Scale bar: 100 μm. (f) Western blots were used to compare VEGFA expression between TUG1 KD group and Nc group. Tubulin expression was detected as the loading control. VEGFA expression was determined as the ratio of densitometric value compared with Tubulin expression. A representative immunoblot was shown along with the quantitative data (n=6; Mann–Whitney U-test; P=0.0112). (g) ELISAs were conducted to compare the difference in plasma VEGFA levels between TUG1 KD group and Nc group (n=6; Mann–Whitney U-test; P=0.0165). *indicated significant difference between Nc group and TUG1 KD group