NF-kappaB p50 promotes tumor cell invasion through negative regulation of invasion suppressor gene CRMP-1 in human lung adenocarcinoma cells.
Lung adenocarcinoma Cl1-5 cells were selected from parental Cl1-0 cells based on their high metastatic potential. In a previous study, CRMP-1, an invasion suppressor gene, was shown to be suppressed in Cl1-5 cells. However, the regulation of CRMP-1 expression has not been explored. In this study, we showed nuclear factor-kappaB controls CRMP-1 expression. The electromobility shift assay showed that while Cl1-0 cells exhibited low NF-kappaB activity in response to TNF-alpha, an abundance of basal and TNF-alpha-induced NF-kappaB-DNA complex was detected in Cl1-5 cells. Supershift-coupled EMSA and Western blotting of nuclear proteins, however, revealed p50 protein, but not classic p65/p50 heterodimer in the complex. ChIP and EMSA demonstrated that p50 binds to a kappaB site residing between -1753 and -1743 of the CRMP-1 promoter region. Transfection of antisense p50 gene into Cl1-5 cells increased the CRMP-1 protein level and decreased the invasive activity of Cl1-5 cells.
Gao
Ming
M
Cancer Research Center, College of Medicine, National Taiwan University, Taipei, Taiwan.
Yeh
Pei Yen
PY
Lu
Yen-Shen
YS
Chang
Wen Ching
WC
Kuo
Min-Liang
ML
Cheng
Ann-Lii
AL
D000230:Adenocarcinoma / Q000235:genetics / Q000473:pathology*; D045744:Cell Line, Tumor; D024202:Electrophoretic Mobility Shift Assay; D015972:Gene Expression Regulation, Neoplastic; D016147:Genes, Tumor Suppressor; D006801:Humans; D008175:Lung Neoplasms / Q000235:genetics / Q000473:pathology*; D052002:NF-kappa B p50 Subunit / Q000378:metabolism*; D009361:Neoplasm Invasiveness; D009419:Nerve Tissue Proteins / Q000235:genetics*; D011401:Promoter Regions, Genetic; D051996:Transcription Factor RelA / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D000098855:Collapsin Response Mediator Protein 1
D052002:NF-kappa B p50 Subunit; D009419:Nerve Tissue Proteins; D051996:Transcription Factor RelA; D014409:Tumor Necrosis Factor-alpha; C102924:CRMP1 protein, human; D000098855:Collapsin Response Mediator Protein 1