Research Paper Volume 13, Issue 7 pp 10672—10687

NET1 promotes HCC growth and metastasis in vitro and in vivo via activating the Akt signaling pathway

MK2206 inhibits Akt phosphorylation and reverses NET1 overexpression-induced HCC phenotype. (A and B) Western blot showing the inhibition of Akt phosphorylation by MK2206 in Hep3B (A) and Huh7 (B) cells with NET1 overexpression. (C and D) Real-time PCR showing the reversal of NET1-induced increased expression of MMP2, Cyclin D1 and Vimentin by MK2206 in Hep3B (C) and Huh7 (D) cells. (E) Crystal violet assays showing the reversal of NET1-induced cell growth by MK2206. Left panel: crystal violet assays. Right panel: measured OD values. (F) Transwell assays showing the reversal of NET1-induced cell migration by MK2206. Left panel: transwell assays. Right panel: calculation of cells that have migrated through the filter following eosin staining. (G) Transwell assays showing the reversal of NET1-induced cell invasion by MK2206. Left panel: transwell assays. Right panel: calculation of cells that have invaded through the filter following eosin staining. All data are presented as mean ± SE. ns not significant. **P ***P ###P

Figure 7. MK2206 inhibits Akt phosphorylation and reverses NET1 overexpression-induced HCC phenotype. (A and B) Western blot showing the inhibition of Akt phosphorylation by MK2206 in Hep3B (A) and Huh7 (B) cells with NET1 overexpression. (C and D) Real-time PCR showing the reversal of NET1-induced increased expression of MMP2, Cyclin D1 and Vimentin by MK2206 in Hep3B (C) and Huh7 (D) cells. (E) Crystal violet assays showing the reversal of NET1-induced cell growth by MK2206. Left panel: crystal violet assays. Right panel: measured OD values. (F) Transwell assays showing the reversal of NET1-induced cell migration by MK2206. Left panel: transwell assays. Right panel: calculation of cells that have migrated through the filter following eosin staining. (G) Transwell assays showing the reversal of NET1-induced cell invasion by MK2206. Left panel: transwell assays. Right panel: calculation of cells that have invaded through the filter following eosin staining. All data are presented as mean ± SE. ns not significant. **P < 0.01, ***P < 0.001 vs vector cells. ###P < 0.001 vs Lenti-NET1 cells.