Advanced Glycation End-Products Enhance Lung Cancer Cell Invasion and Migration

Abstract

Effects of carboxymethyllysine (CML) and pentosidine, two advanced glycation end-products (AGEs), upon invasion and migration in A549 and Calu-6 cells, two non-small cell lung cancer (NSCLC) cell lines were examined. CML or pentosidine at 1, 2, 4, 8 or 16 μmol/L were added into cells. Proliferation, invasion and migration were measured. CML or pentosidine at 4-16 μmol/L promoted invasion and migration in both cell lines, and increased the production of reactive oxygen species, tumor necrosis factor-α, interleukin-6 and transforming growth factor-β1. CML or pentosidine at 2-16 μmol/L up-regulated the protein expression of AGE receptor, p47(phox), intercellular adhesion molecule-1 and fibronectin in test NSCLC cells. Matrix metalloproteinase-2 protein expression in A549 and Calu-6 cells was increased by CML or pentosidine at 4-16 μmol/L. These two AGEs at 2-16 μmol/L enhanced nuclear factor κ-B (NF-κ B) p65 protein expression and p38 phosphorylation in A549 cells. However, CML or pentosidine at 4-16 μmol/L up-regulated NF-κB p65 and p-p38 protein expression in Calu-6 cells. These findings suggest that CML and pentosidine, by promoting the invasion, migration and production of associated factors, benefit NSCLC metastasis.

Keywords: CML; invasion; migration; non-small cell lung cancer; pentosidine.

Figure 1

Effects of CML or pentosidine at 0 (control), 1, 2, 4, 8 or 16 μmol/L upon protein expression of Bcl-2, Bax, caspase-3 and caspase-8 in human A549 and Calu-6 cells. Cells were exposed to CML or pentosidine for 18 h at 37 °C. Data are mean ± SD (n = 10).

Figure 2

Effects of CML or pentosidine at 0 (control), 1, 2, 4, 8 or 16 μmol/L upon protein expression of RAGE, p47^phox and gp91^phox in human A549 and Calu-6 cells. Cells were exposed to CML or pentosidine for 18 h at 37 °C. Data are mean ± SD (n = 10) and shown in the following table. ^a–d Means within a column without a common letter differ, p < 0.05.

Figure 3

Effects of CML or pentosidine at 0 (control), 1, 2, 4, 8 or 16 μmol/L upon protein expression of VEGF, ICAM-1, fibronectin, MMP-2 and MMP-9 in human A549 and Calu-6 cells. Cells were exposed to CML or pentosidine for 18 h at 37 °C. Data are mean ± SD (n = 10) and shown in the following table. ^a–e Means within a column without a common letter differ, p < 0.05.

Figure 4

Effects of CML or pentosidine at 0 (control), 1, 2, 4, 8 or 16 μmol/L upon protein expression of NF-κB and MAPK in human A549 and Calu-6 cells. Cells were exposed to CML or pentosidine for 18 h at 37 °C. Data are mean ± SD (n = 10), and shown in the following table. ^a–e Means within a column without a common letter differ, p < 0.05.

Figure 5

Effects of CML or pentosidine at 0 (control), 1, 2, 4, 8 or 16 μmol/L upon NF-κB p50/65 DNA binding activity, determined as OD450 nm/mg protein, in human A549 and Calu-6 cells. Cells were exposed to CML or pentosidine for 18 h at 37 °C. Data are mean ± SD (n = 10). ^a–e Means among bars without a common letter differ, p < 0.05.