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.
Zhang
Jian
J
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
Xu
Yongjian
Y
Xiong
Weining
W
Zhang
Zhenxiang
Z
Du
Chunling
C
Qiao
Lifen
L
Ni
Wang
W
Chen
Shixin
S
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