Figure 6
Low frequency magnetic fields induce lung cancer cell iron metabolism dysfunction. A549 and LLC cells were treated with MF or Sham MF for 2–4 days. (A) Cells were washed, digested with 5% HNO3 and the supernatant collected for intracellular non-haem iron estimation using flame atomic absorption spectrometer. (B) The mRNA levels of TfR and ferritin in LLC cells were detected using Q-PCR. (C) Protein level of TfR and ferritin in LLC cells were detected using western blot. Numbers under each blot are relative intensity of the blot. (D) Surface expression of TfR on LLC cells was detected using flow cytometry. (E) Immunofluorescence analysis of A549 and LLC cells treated with MF or Sham MF for 2days. Fluorescent probe Phen Green SK (PG-SK) was used for monitoring labile iron pool (Green). Cells treated with 100 μM ferrous sulfate (FeSO4) for 10 min were taken as positive control. Cells incubated with 100 μM DFO for 15 min were taken as negative control. Scale bars, 20 µm. (F) Immunofluorescence analysis of A549 cells treated by MF or Sham MF for 2 days. The cells were fixed and stained with ferritin antibody (red), PG-SK (Green), and DAPI (blue) respectively. Scale bars, 20 µm. (G) Measurement of non-haem iron content in tumor tissue of LLC murine model. (n = 6) (H) Immunochemistry analyzes TfR and ferritin-H expression in tumor tissue of LLC murine model. (n = 6); Scale bars, 100 µm. (I) IHC images were calculated using Image Pro Plus software 6.0. All experiments were repeated three times. Data represent Mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001.
