Fig. 2

MACC1-AS1 is induced under metabolic stress and promotes GC progression. a-b MACC1-AS1-overexpressing or vector-transfected MKN45 cells were injected into the caudal vein of nude mice (n = 5, 1 × 10⁶ per mouse). Gross lung metastatic tumors (a) and H&E stained sections of lungs showed that overexpression of MACC1-AS1 promoted the metastasis of GC. Immunohistochemical staining showed that the MACC1-AS1 overexpression group exhibited higher MACC1 expression with higher rates of proliferation, as measured by Ki-67 staining, and lower numbers of oxidative lesions, based on 8-OHdG staining (b). c Quantitative analysis of lung metastatic nodules showed that the number of nodules were not significantly different, while MACC1-AS1 overexpression group exhibited larger relative area of nodules of the lung. d Quantitative analysis of IHC staining showing MACC1-AS1 overexpression group exhibited higher expression of MACC1, Ki-67 but lower expression of 8-OHdG. e-g qPCR results showing that MACC1-AS1 was increased after glucose deprivation (e), 2-DG (10 mM) (f) or H₂O₂ (100 μM) (g) treatment. *P < 0.05, ***P < 0.001. h qPCR results showing that glucose deprivation-derived MACC1-AS1 upregulation was reversed by NAC treatment (8 mM). **P < 0.01. i qPCR results showing that MACC1-AS1 was upregulated under culture conditions of sustained low glucose for 1 month (1, 0.5 g/L) compared to normal glucose (2 g/L). *P < 0.05, ***P < 0.001. j Western blotting results showing that GLUT1 and HK2 were increased under culture conditions of sustained low glucose for 1 month. k-l Representative staining and quantitative analysis of lung metastatic nodules showing MACC1-AS1 overexpression group exhibited higher expression of GLUT1, HK2 and LDH