Inhibitory activity of nuclear factor-kappaB potentiates cisplatin-induced apoptosis in A549 cells.
Whether inhibiting the activity of nuclear factor (NF)-kappaB potentiates cisplatin-induced apoptosis in non-small cell lung cell line A549 cells was investigated. The recombinant plasmid pcDNA3.1 (+)/IkappaBalpha expressing IkappaBalpha was constructed. The in vitro cultured A549 cells were transfected with pcDNA3.1 (+)/IkappaBalpha alone, or pcDNA3.1 (+)/IkappaBalpha combined with cisplatin. The mitochondrial membrane potential (Deltapsim) was determined by rhodamine 123, the activity of caspase-3 was tested by colorimetric assay, and cell apoptosis was detected by flow cytometry with the annexin V /propidium iodide assay. The results showed that the activity of NF-kappaB in A549 cells was inhibited by transfecting pcDNA3.1(+)/IkappaBalpha. Transfection of pcDNA3.1(+)/IkappaBalpha alone did not promote apoptosis. Treatment of cisplatin alone had a little effect on cell apoptosis. Transfection of pcDNA3.1(+)/IkappaBalpha combined with cisplatin treatment significantly induced apoptosis of A549 cells. It was concluded that inhibiting the activity of NF-B potentiated cisplatin-induced apoptosis of A549 cells.
Department of Respiratory Medicine, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Key Laboratory of Pulmonary Diseases of Ministry of Health of China, Wuhan 430030, China. zhangjiandoc@126.com
Jian
J
Zhang
Yongjian
Y
Xu
Weining
W
Xiong
Zhenxiang
Z
Zhang
Chunling
C
Du
Lifen
L
Qiao
Wang
W
Ni
Shixin
S
Chen
D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis; D053148:Caspase 3 / Q000378:metabolism; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000378:metabolism; D002945:Cisplatin / Q000494:pharmacology*; D005456:Fluorescent Dyes / Q000494:pharmacology; D006801:Humans; D020672:I-kappa B Proteins / Q000378:metabolism; D008564:Membrane Potentials; D008928:Mitochondria / Q000378:metabolism; D000072000:NF-KappaB Inhibitor alpha; D016328:NF-kappa B / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D010957:Plasmids / Q000378:metabolism; D020112:Rhodamine 123 / Q000494:pharmacology; D014162:Transfection
D000970:Antineoplastic Agents; D005456:Fluorescent Dyes; D020672:I-kappa B Proteins; D016328:NF-kappa B; C000606204:NFKBIA protein, human; D000072000:NF-KappaB Inhibitor alpha; D020112:Rhodamine 123; D053148:Caspase 3; D002945:Cisplatin