Figure 7.

NHEG1 Exerts Oncogenic Roles via Interacting with DDX5

(A) Western blot assay showing the expression of DDX5 in SH-SY5Y and SK-N-SH cells stably transfected with empty vector (Mock) or NHEG1 and those cotransfected with scramble shRNA (sh-Scb) or sh-DDX5-1. (B–D) Quantification of flow cytometric (B), soft agar (C), and Transwell Matrigel invasion (D) assays indicating the cell-cycle progression, anchorage-independent growth, and invasion capability of NB cells stably transfected with mock or NHEG1 and those cotransfected with sh-Scb or sh-DDX5-1 (n = 6). (E–H) Representative images (E, top panel), in vivo growth curve (E, bottom panel), tumor weight (F), and Ki-67 positive rate (G) at the end points of xenograft tumors in athymic nude mice formed by hypodermic injection of SH-SY5Y cells stably transfected with mock, NHEG1, sh-Scb, and sh-DDX5-1 (n = 5 for each group). Representative images (E, top panel), H&E staining (E, arrowheads), and quantification (H) of lung metastatic colonies in athymic nude mice (n = 5 for each group) after tail-vein injection of SH-SY5Y cells stably transfected with mock, NHEG1, sh-Scb, or sh-DDX5-1. Scale bars, 100 μm. (I) Kaplan-Meier curves of nude mice treated with tail-vein injection of SH-SY5Y cells stably transfected with mock or NHEG1 and those cotransfected with sh-Scb or sh-DDX5-1 (n = 5 for each group). ANOVA analyzed the difference in (B)–(H). Log-rank test for survival comparison in (I). *p < 0.01 versus mock + sh-Scb. NS, not significant. Data are shown as mean ± SEM (error bars) and representative of three independent experiments in (A)–(D).