The role of constitutive NF-κB activity in PC-3 human prostate cancer cell invasive behavior
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The purpose of this study was to determine if increased NF-κB activity of highly invasive PC-3 cells contributed to their invasive behavior. Increased NF-κB activity has been observed in several malignant tumors and it may have an important role in tumorigenesis, progression and chemotherapy resistance. By serial selection, we obtained invasion variant PC-3 cell sublines. The PC-3 High Invasive cells invade readily through a Matrigel® reconstituted basement membrane while PC-3 Low Invasive cells have low baseline invasion activity. In these studies, we discovered that NF-κB DNA binding activity was increased in PC-3 High Invasive cells when compared to PC-3 Low Invasive cells by electrophoretic mobility shift assay (EMSA). Gel supershift assays showed a 4-fold increase in p65 containing complexes and a 2.2-fold increase in the p50 containing complexes in the PC-3 High Invasive cells. Luciferase reporter assays showed that NF-κB dependent transcription activity was increased 10.2±2.5-fold in the highly invasive cells (P<0.002). The PC-3 High Invasive cells showed a constitutive increase in phospho-IκBα and introduction of the super-repressor IκBα S32/36A inhibited NF-κB activity to 19.2±2.5 percent of control transfected cells (P≤0.001). The IκBα super-repressor reduced the basement membrane invasion of PC-3 High Invasive cells from 6.2±1.1 to 3.8±0.4 percent (P<0.002) with no decrease in cell viability or proliferation. These results demonstrate that increased NF-κB activity contributed directly to the invasive behavior of PC-3 High Invasive prostate cancer cells.
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- The role of constitutive NF-κB activity in PC-3 human prostate cancer cell invasive behavior
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