RRC ID 38555
著者 Ikari A, Sato T, Takiguchi A, Atomi K, Yamazaki Y, Sugatani J.
タイトル Claudin-2 knockdown decreases matrix metalloproteinase-9 activity and cell migration via suppression of nuclear Sp1 in A549 cells.
ジャーナル Life Sci
Abstract AIMS:Claudin expression is altered in lung cancer, but the pathophysiological role of claudin is not well understood. We examined the effect of claudin-2 expression on cell migration using human adenocarcinoma A549 cells.
MAIN METHODS:The mRNA level was measured by real time polymerase chain reaction. To knockdown claudin-2 expression, we made the cells expressing doxycycline-inducible claudin-2 shRNA vector. The protein level was examined by Western blotting. Cell migration was measured by wound-healing assay. The enzymatic activity of MMP-9 was assessed by gelatin zymography.
KEY FINDINGS:In A549 cells, claudin-2 expression was higher than in normal lung tissue. Claudin-2 knockdown did not affect the expression of other junctional proteins including claudin-1, occludin and E-cadherin. Claudin-2 knockdown decreased cell migration concomitant with a decrease in the mRNA level and enzymatic activity of MMP-9. The expression level of Sp1 in the nuclei was decreased by claudin-2 knockdown. In contrast, the expression levels of c-Fos, c-Jun and NF-kB p65 in the nuclei were not changed by claudin-2 knockdown. The knockdown of Sp1 expression by siRNA decreased cell migration, and the mRNA expression, enzymatic activity, and promoter activity of MMP-9.
SIGNIFICANCE:Claudin-2 may increase the mRNA level and enzymatic activity of MMP-9 mediated by the elevation of nuclear distribution of Sp1, resulting in the up-regulation of A549 cell migration.
巻・号 88(13-14)
ページ 628-33
公開日 2011-3-28
DOI 10.1016/j.lfs.2011.02.002
PII S0024-3205(11)00063-4
PMID 21315735
MeSH Cell Line, Tumor Cell Movement* / genetics Cell Nucleus / enzymology Cell Nucleus / genetics Cell Nucleus / metabolism* Claudins Cytoplasm / enzymology Cytoplasm / genetics Cytoplasm / metabolism Electrophoresis, Polyacrylamide Gel Gene Knockdown Techniques Humans Lung / enzymology Lung / metabolism Lung Neoplasms / enzymology Lung Neoplasms / genetics Lung Neoplasms / metabolism* Lung Neoplasms / pathology Matrix Metalloproteinase 9 / metabolism* Membrane Proteins / biosynthesis* Membrane Proteins / genetics Microscopy, Confocal Reverse Transcriptase Polymerase Chain Reaction Sp1 Transcription Factor / antagonists & inhibitors* Sp1 Transcription Factor / genetics Transfection
IF 3.647
引用数 35
WOS 分野 PHARMACOLOGY & PHARMACY MEDICINE, RESEARCH & EXPERIMENTAL
リソース情報
ヒト・動物細胞 A549(RCB0098)