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. 2017 Feb 2;8(26):42087–42097. doi: 10.18632/oncotarget.14992

Figure 2. CRNDE knockdown inhibits tumor growth and tumor angiogenesis in vivo.

Figure 2

An in vivo hepatoblastoma model was established by injection of CRNDE knockdown HuH-6 cells and scrambled shRNA-transfected HuH-6 cells into the left flank of nude mice (C57BL/6). CRNDE expression in the tumors isolated from CRNDE knockdown (KD) group and negative control group (Scr) were detected by qRT-PCRs after 4-week tumor inoculation (A; Student's t-test; P=0.0023). The volume of solid tumor was measured every week using a vernier caliper [B; n=6; Mann-Whitney U-test; P=0.0285 (2 w), P=0.0209 (3 w), P=0.0134 (4 w)]. The tumors were weighed immediately after 4-week tumor inoculation (C; Mann-Whitney U-test; P=0.0011). CD31 and PCNA staining was performed to detect microvascular density D. and cell proliferation F. in the hepatoblastoma tissues after 4-week tumor inoculation. Vessels with a clearly defined lumen or well-defined linear vessel shape but not single endothelial cells were considered for microvessel density assessment. A representative image and statistical result was shown [n=10 slices; Mann-Whitney U-test; P=0.0034 (D) and P=0.0022 (F)]. Scale bar: 100 μm. ELISA assays were conducted to detect CD31 expression after 4-week tumor inoculation (E, n=5; Mann-Whitney U-test; P=0.0024). ELISAs were conducted to compare VEGFA G. and Ang-2 H. levels between hepatoblastoma tissue of CRNDE KD group and Scr group after 4-week tumor inoculation (n=6; Mann-Whitney U-test; P=0.0124). “*” indicated significant difference between Scr group and CRNDE KD group.