Figure 5.

Inhibition of growth and molecular effects of MET and IR in human LC xenografts. (A) Effects on xenograft growth. Twenty-four and sixteen male Balb/c-nude mice were grafted into the right flank with 1 × 10⁶ A549 or H1299 cells respectively. Treatment with MET, IR or combination was initiated once tumour volume reached 100 mm³. Tumour volume was measured every 5 days for a period of 8 weeks, as described in Materials and Methods. Average tumour volume data of four or six animals per treatment group are shown (mean±s.e.). Final tumour volumes were: A549: control: 679.5±50.6 mm³, MET: 470.53±35.2 mm³, IR: 342.9±21.8 mm³ and MET+IR: 257.9±15.2 mm³ and H1299: control: 1129.39±39.17 mm³, MET: 646.02±40.43 mm³, IR: 428.02±18.89 mm³ and MET+IR: 300.25±24.25 mm³. Statistically significant differences between the indicated treatment groups (*P<0.05 and **P<0.001). (B) Effects of MET and IR treatments on expression and activation of molecular markers. Tumours collected from control (CON), MET, IR and MET+IR treatment groups of A549 cohort were subjected to lysis and immunoblotting with antibodies against the indicated markers. Representative immunoblots of three independent experiments are shown. (C) Immunohistochemical analysis of P-AMPK expression. Analysis of A549 tumours from four different treatment groups using an anti-P-AMPKα (Thr172) antibody. Representative images from three independent experiments are shown. (D) Densitometric analysis of immunoblotting experiments (n=6 in each group). Data are presented as mean±s.e. of normalised densitometry values. *P⩽0.05; **P⩽0.001 statistically significant difference between treatment groups and control for each marker.