The role of constitutive NF-κB activity in PC-3 human prostate cancer cell invasive behavior
The role of constitutive NF-κB activity in PC-3 human prostate cancer cell invasive behavior
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DOI：
10.1023/a:1011845725394
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发表时间：
2001-01-01
影响因子：
4
通讯作者：
Kajdacsy-Balla, A
中科院分区：
文献类型：
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作者：
Lindholm, PF;Bub, J;Kajdacsy-Balla, A
本研究的目的是确定高侵袭性PC-3细胞核因子-kappaB活性增加是否与其侵袭行为有关。在多种恶性肿瘤中已观察到核因子-kappaB活性升高，它可能在肿瘤的发生、发展和化疗耐药中起重要作用。通过连续筛选，获得了侵袭力不同的PC-3细胞亚系。PC-3高侵袭性细胞很容易通过Matrigel(R)重组基底膜侵袭，而PC-3低侵袭性细胞具有低基线侵袭活性。在这些研究中，我们发现与PC-3低侵袭性细胞相比，高侵袭性PC-3细胞中的NF-kappaB DNA结合活性增加。凝胶超迁移实验显示PC-3高侵袭性细胞中含有p65的复合体增加了4倍，含有p50的复合体增加了2.2倍。荧光素酶报告分析显示，在高侵袭性细胞中，依赖于NF-kappaB的转录活性增加了10.2+/-2.5倍(P<；0.002)。PC-3高侵袭性细胞表现出磷酸化I kappaBα的结构性增加，而超阻遏因子I kappaBαS32/36A的引入抑制了19.2+/-2.5%的对照细胞(P<或等于0.001)。I kappaBα超抑制物可使PC-3高侵袭性细胞基底膜侵袭能力从6.2%+/-1.1%降至3.8%+/-0.4%(P<；0.002)，细胞存活率和增殖能力未见下降。这些结果表明，核因子-kappaB活性的增加直接参与了PC-3高侵袭性前列腺癌的侵袭行为。
The purpose of this study was to determine if increased NF-kappaB activity of highly invasive PC-3 cells contributed to their invasive behavior. Increased NF-kappaB 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(R) reconstituted basement membrane while PC-3 Low Invasive cells have low baseline invasion activity. In these studies, we discovered that NF-kappaB 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-kappaB 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-I kappaB alpha and introduction of the super-repressor I kappaB alpha S32/36A inhibited NF-kappaB activity to 19.2 +/- 2.5 percent of control transfected cells (P less than or equal to 0.001). The I kappaB alpha 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-kappaB activity contributed directly to the invasive behavior of PC-3 High Invasive prostate cancer cells.