Oncogenic potential of NM23-H2 in HLK3 cells. (A) NM23-H2-expressing HLK3 cells (HNM10 and HNM14) but not vector control cells (HVC3 and HVC6) grow as tumors in nude mice. Tumors are shown 58 days post-injection. (B) Growth of the tumor mass in NM23-H2 expressing (HNM10 and HNM14) and vector control (HVC3 and HVC6) cells injected into nude mice. NM23-H2 expressing cell lines grew faster than control cells and formed tumor masses. The tumor volume was measured as a function of time (the animal was finally euthanized due to an ulceration of the primary tumor). Each value represents the mean ± SE. (C) Histology of NM23-H2-derived tumors stained with NM23-H2 (left panels) and hepatocyte (right panels) antibodies. HCC tissue staining was used as a positive control (original magnification, 200×). (D) Western blot analysis showing c-Myc, cyclin D1 (CCND1), and p65 over-expression in stable cell lines (HNM10 and HNM14), xenotransplanted tumors (mHNM10 and mHNM14), parent cells (HLK3), and vector control cells (HVC3 and HVC6). (E) Immunofluorescence assay showing that ectopic NM23-H2 expression overlaps with p65 (NF-κB), CCND1, and c-Myc over-expression in HLK3 cells. Hoechst 33258 staining of nuclei (blue). Immunoreactivity of NM23-H2 and Flag (FITC) is merged with c-Myc, CCND1, and p65 (TRITC); bar, 20 µm.