Figure 5.

Downregulation of miR-362-5p inhibits the SW780 xenograft tumor growth. (A) A hairpin sequence containing the 100% complementary nucleotide sequence of miR-362-5p was constructed into pRNA-H1.1/Adeno vector. The SW780 cells were transfected with vector containing anti-miR-362-5p (2 μg) or miR-NC. Then transfected SW780 cells were selected using 300 µg/ml G418. The Balb/c nude mice were injected subcutaneously into the left flank with 1×10⁷ SW780 cells stable transfected cells (100 μl PBS) to form xenograft tumors. The tumor volumes were measured on day 25. (B) The relative expression of miR-362-5p in xenograft tumor tissues from anti-miR-362-5p or miR-NC injected mice were analyzed by qRT-PCR. The expression was displayed as fold of NC. (C) The tumor growth curves were determined by assessing the tumor volumes every 3 days from day 7 to day 25 by measuring two perpendicular diameters and the tumor volumes were calculated according to the following formula: [(width)² × length]/2. (D) The xenograft tumor weight was measured on day 25. (E) The expression levels of Ki-67 in the xenograft tumor tissues were determined using immunohistochemical assay. (F) The protein levels of QKI, Cyclin D, MacroH2A1.1, MacroH2A1.2, p27, c-Fos, PARP-1, and E2F1 in xenograft tumor tissues were detected by western blot. GAPDH was used as an internal control in western blot. *p < 0.05 vs. NC controls.