[Inhibition of NF-kappaB through IkappaBalpha transfection affects invasion of human lung cancer cell line A549].
OBJECTIVE
Recent studies have shown that activation of nuclear factor-kappaB(NF-kappaB) can regulate the invasion and metastasis of cancer cells. The present study was to investigate inhibition of NF-kappaB activity on invasion of human lung cancer cell line A549 and the possible mechanism.


METHODS
The recombinant plasmid pcDNA3.1(+)/IkappaBalpha, expressing the alpha isoform (IkappaBalpha) of the NF-kappaB inhibitor, was constructed. A549 cells were cultured in vitro and divided into non-transfection group, pcDNA3.1(+) transfected group and pcDNA3.1(+)/IkappaBalpha transfected group. The expression of IkappaBalpha was detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting. The activity of NF-kappaB was determined by electrophoretic mobility shift assay (EMSA). Invasion of A549 cells was assessed by transwell chamber assay. The expression of MMP-2 and MMP-9 was detected by RT-PCR and gelatin zymography.


RESULTS
Plasmid pcDNA3.1(+)/IkappaBalpha was successfully constructed and expressed in A549 cells. The activity of NF-kappaB, the number of invasive cells, the activity of MMP-2 and MMP-9 of A549 cells in pcDNA3.1(+)/IkappaBalpha transfected group were significantly lower than those in non-transfection group and pcDNA3.1(+) transfected group (all P<0.05).


CONCLUSIONS
Transfection of IkappaBalpha can inhibit NF-kappaB activity, thus inhibit cell invasion of A549, which may be through the down-regulation of MMP-2 and MMP-9 expressions.
Zhang
Jian
J
Department of Respiratory Medicine, Tongji Hospital, Tongji Medical School, Huazhong University of Science and Technology,Wuhan, Hubei, 430030, P R China.
Xu
Yong-Jian
YJ
Xiong
Wei-Ning
WN
Zhang
Zhen-Xiang
ZX
DU
Chun-Ling
CL
Qiao
Li-Fen
LF
Ni
Wang
W
Chen
Shi-Xin
SX
D045744:Cell Line, Tumor; D006801:Humans; D020672:I-kappa B Proteins / Q000235:genetics / Q000502:physiology*; D008175:Lung Neoplasms / Q000473:pathology*; D020778:Matrix Metalloproteinase 2 / Q000032:analysis / Q000235:genetics; D020780:Matrix Metalloproteinase 9 / Q000032:analysis / Q000235:genetics; D061965:Matrix Metalloproteinase Inhibitors; D000072000:NF-KappaB Inhibitor alpha; D016328:NF-kappa B / Q000037:antagonists & inhibitors*; D009361:Neoplasm Invasiveness; D012333:RNA, Messenger / Q000032:analysis; D014162:Transfection
D020672:I-kappa B Proteins; D061965:Matrix Metalloproteinase Inhibitors; D016328:NF-kappa B; C000606204:NFKBIA protein, human; D012333:RNA, Messenger; D000072000:NF-KappaB Inhibitor alpha; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9