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標題Title: Anti-sense morpholino oligonucleotide assay shows critical involvement for NF-jB activation in the production of Wnt-1 protein by HepG2 cells: oncology implications
作者Authors: 溫義輝,Chi-Shu Sun..等
上傳單位Department: 生物科技系
上傳時間Date: 2009-11-16
上傳者Author: 溫義輝
審核單位Department: 生物科技系
審核老師Teacher: 溫義輝
檔案類型Categories: 論文Thesis
關鍵詞Keyword: anti-sense strategy, chromatin immunoprecipitation assay, HepG2 cells, Ikappa B, Morpholino oligonucleotide, NF-kappaB, Wnt-1 protein
摘要Abstract: The link of proto-oncogenic protein Wnt-1
production with NF-kappaB activation has been functionally
demonstrated in PC12 cells, a rat pheochromocytoma cell
line of neural crest lineage, while it is not yet verified in
human cells. The link can be indirectly supported in our
previous report that functional proteomics identifies
enhanced expression of NF-kappaB-associated Wnt-1
production in human hepatocellular carcinoma tissues. This
study aimed to further validate this link in human cells
using anti-sense strategy. The effects of sequence-specific
anti-sense morpholino oligonucleotides (ONs) targeting
against pre-mRNA sequences of human p50 and p65 subunits
of NF-kappaB as well as Wnt-1 genes were investigated. It
revealed that all the three morpholino ONs inhibited NF-kappaB
activation in human hepatoblastoma cell line HepG2
cells along with decreased Wnt-1 production. Chromatin
immunoprecipitation assay ascertained the direct binding
of NF-kappaB-p50 to the Wnt-1 promoter. Additionally, anti-
P50 and anti-P65 morpholino ONs also repressed the
phosphorylation of IkappaBa which temporarily correlated
with the inhibition of NF-kappaB activation accompanied by
decreased Wnt-1 production by HepG2 cells. In summary,
NF-kappaB activation is critically involved in the production of
Wnt-1 by HepG2 cells. These results may have important
oncology implications in treating patients with NF-kappaBassociated
Wnt-1-producing cancers.

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