Down-Regulation of eIF4GII by miR-520c-3p Represses Diffuse Large B Cell Lymphoma Development
Figure 6
Overexpression of miR-520c-3p in DLBCL diminished colony formation in clonogenic assay and decreased tumor growth in xenograft model.
(A) Farage cells after transfection with either Pre-miR-Ctrl or Pre-miR-520c-3p were cultured in agarose/medium for two weeks. Representative pictures (10×) are shown. Graphs represent the means and SD of three independent experiments. (B) Farage cells were transfected with Ctrl siRNA or eIF4GII siRNA, assay was performed and analyzed as described in (A). * p<0.05. (C and D) SCID Beige mice (n = 5) received a subcutaneous injection of SUDHL4 cells either expressing empty pCDH-Vector (on left sites; red arrow) or overexpressing miR-520c-3p, pCDH-520c-3p (on right sites; green arrow). The average tumor volume of each group with SEM is shown as a function of time. The repeated measure ANOVA showed a significant effect of time on tumors growth F(8,64) = 40.23, p<0.001, and significant repression of growth by miR-520c-3p as revealed by significant effect of treatment F(1,8) = 49.98, p<0.001 and significant treatment x time interaction F(8,64) = 5.93, p<0.001. (E) Lysates from SUDHL4 xenograft tumors obtained in C and D were fractionated by centrifugation through 10–50% linear sucrose gradients, and the polysome profiles were studied. (F) Protein extracts from xenograft tumors from (C) and (D) were subjected to Western blot analysis using indicated senescence markers antibodies.