Fig. 8.

Forced expression of furin promotes EMT and metastatic potential of PTC cells. (A) TPC‐1 cells were transfected with either an empty vector or FURIN cDNA. After cell lysis, proteins were immuno‐blotted with antibodies against Furin, E‐cadherin, N‐cadherin, Zeb1, Twist, and β‐Actin as indicated (n = 3). (B, C) Forced expression of furin increases invasion. TPC‐1 cells were transfected with either an empty vector or FURIN cDNA. Cells were seeded into the upper compartment of invasion chambers. The bottom chambers were filled with RPMI media. After 24 h incubation, invaded cells were fixed, stained, and quantified (scale bar = 1 mm). Data were presented as mean ± SD (n = 3). (D) Forced expression of furin increases migration. After transfection, cells were seeded into the upper compartment of migration chambers. The bottom chambers were filled with RPMI media. After 24 h incubation, migrated cells were fixed, stained, and quantified. Data were presented as mean ± SD (n = 3). (E) Inhibition of MEK reduces EMT in furin‐expressing cells. TPC‐1 cells carrying either empty vector or FURIN cDNA were treated with selumetinib (20 nM) for 48 h. After cell lysis, proteins were immuno‐blotted with antibodies against Furin, E‐cadherin, N‐cadherin, Zeb1, Twist, and β‐Actin as indicated (n = 3). (F, G) Selumetinib decreases the invasion and migration of Furin expressing cells. TPC‐1 cells carrying either empty vector or FURIN cDNA were treated with selumetinib (20 nM) for 48 h and the cells were subjected to invasion and migration assay. Data were presented as mean ± SD (n = 3). Statistical analyses were performed using two‐tailed Student's t‐tests. *P < 0.05.