Fig. 4.

hENT1-dependent growth of high-grade meningioma cells. HKBMM and IOMM-Lee cells transfected twice with siControl, sihENT1 (#1 or #2), or sidCK (#1 or #2) were harvested 3 days after the second transfection and subjected to Western blotting (A and B) and cell cycle analyses (n = 3-5) (E and F). Alternatively, transfected cells were seeded on 96-well plates (1000 cells/well for HKBMM and 500 cells/well for IOMM-Lee) on the day after the second transfection and, after 7 days, cell viability was evaluated by the WST-8 assay (n = 5-7) (C and D). (G) IOMM-Lee cells transfected twice with siControl, sihENT1#1, or sidCK#2 siRNAs were subcutaneously implanted into nude mice (1 × 10⁶ viable cells/injection, n = 8, each group) on the day following the second transfection, and the tumor volume was measured. In (C) through (G), values in the graphs were the mean and SD. In (C) and (D), the average absorbance for siControl was set to 100% for each experiment, and P values were calculated by a one-sample t-test. In (E) and (F), P values from comparisons with the values of siControl were calculated by a one-way ANOVA with Tukey’s post-hoc test. In (G), P values from comparisons with the values of siControl group were calculated by the Kruskal-Wallis test. *P < .05. Abbreviations: ANOVA, analysis of variance; dCK, deoxycytidine kinase; hENT1, human equilibrative nucleoside transporter 1.