Figure 4

Inflammation results in sustained oxidative damage to DNA. A. The levels of the DNA damage marker γ-H2AX^(S139) and the metabolic regulator SIRT1 are remarkably affected by treatment with CM and H₂O₂ in the absence of LNAC. B. SIRT1 mRNA expression in LNCaP cells is altered by treatment with CM (including 125 and 250 pg/ml of TNFα) and 250 μM H₂O₂ with and without 10 mM LNAC. C. Caspase 3 and p-ATM^(S1981) levels remain lower in the treated cells in comparison to controls, indicating that the cells failed to activate apoptosis and the DNA damage response upon oxidative DNA damage. LPS (10 ng/ml) treatment was used as positive control for caspase-3 cleavage and activation. D. LNCaP cells were exposed to CM (including 50 and 100 pg/ml of TNFα) and H₂O₂ (50, 100 and 200 μM) for 2 weeks. Chronic exposure to CM, but not H₂O₂, results in the loss of NKX3.1. E. Similar to the acute treatments (24 h), chronic CM exposure (100 pg/ml of TNFα) also results in increases in QSCN6 and GPX2 expression but a decrease in GPX3 expression in LNCaP cells. Red bars represent upregulation, green bars represent downregulation. F. Chronic CM exposure, but not treatment with H₂O₂, results in an increase in the intracellular ROS level; chronic exposure to H₂O₂ does not affect the level of ROS in LNCaP cells. The ROS level was measured using a DCFH-DA assay with four replicates.