Zhang Jian, Xu Yongjian, Xiong Weining, Zhang Zhenxiang, Du Chunling, Qiao Lifen, Ni Wang, Chen Shixin
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.
J Huazhong Univ Sci Technolog Med Sci. 2008 Jun;28(3):251-6. doi: 10.1007/s11596-008-0305-4. Epub 2008 Jun 19.
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.
研究了抑制核因子(NF)-κB的活性是否能增强顺铂诱导的非小细胞肺癌细胞系A549细胞凋亡。构建了表达IκBα的重组质粒pcDNA3.1(+)/IκBα。将体外培养的A549细胞单独用pcDNA3.1(+)/IκBα转染,或用pcDNA3.1(+)/IκBα与顺铂联合转染。用罗丹明123测定线粒体膜电位(Δψm),用比色法检测caspase-3的活性,并用膜联蛋白V/碘化丙啶法通过流式细胞术检测细胞凋亡。结果表明,转染pcDNA3.1(+)/IκBα可抑制A549细胞中NF-κB的活性。单独转染pcDNA3.1(+)/IκBα不促进细胞凋亡。单独用顺铂处理对细胞凋亡影响较小。转染pcDNA3.1(+)/IκBα并联合顺铂处理可显著诱导A549细胞凋亡。结论是,抑制NF-κB的活性可增强顺铂诱导的A549细胞凋亡。