Figure 6 | Cell Death & Disease

Figure 6

From: Cancer-derived immunoglobulin G promotes tumor cell growth and proliferation through inducing production of reactive oxygen species

Figure 6

ROS regulated the growth and proliferation of IgG-deficient cancer cells. (AD) HeLa cells were transfected with IGHG1 siRNA or scrambled siRNA for 72 h, and then stimulated with different concentrations of scavengers including CAT (0, 250, 500 and 1000 μg/ml), DMSO (0, 1, 10 and 20%), NAC (0, 5, 10 and 20 mM), SOD (0, 500, 1000 and 2000 U/ml) for 48 h. Cell viability was analyzed with a cell proliferation assay. (E) A portion of IgG-deficient cells were pretreated for 1 h with H2O2 (2 μM) and then treated with or without CAT (500 μg/ml), DMSO (1%), NAC (20 mM) and SOD (2000 U/ml) for 48 h. Another portion was directly stimulated with the above scavengers. Cell proliferation was determined with MTS. Results are presented as percentage of cell proliferation in comparison with the negative control. Data shown are the mean±S.D. of three independent experiments (*P<0.05; **P<0.01). Similar experiments were performed for HEp-2 and PC3 cells (F and G). (H and I) A portion of IgG-deficient cells was pretreated for 1 h with H2O2 (2 μM) and then treated with or without CAT (500 μg/ml) or SOD (2000 U/ml) for 48 h. Another portion was directly stimulated with CAT or SOD. Cells were then incubated with fluorescent probe DCFH-DA or DHE at a final concentration of 10 μM in high-glucose DMEM without FBS for 20 min at 37 °C, washed three times with serum-free medium and then were observed with a confocal microscope. Scale bar, 50 μM. (J and K) A portion of IgG-deficient cells were pretreated for 1 h with H2O2 (2 μM) and then treated with or without CAT (500 μg/ml), DMSO (1%), NAC (20 mM), SOD (2000 U/ml) for 48 h. Another portion was directly stimulated with the above scavengers. Whole-cell lysates were fractionated by SDS-PAGE and blotted with anti-p-ERK and anti-IgGγ antibodies

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