The Changes Of Nuclear Transcription Factor Nf-kb And Ap-1 After The Complements Of The Bujingjiedu Treat On Lung Cancer A549 Cells

Essence deficiency and poison are important lung cancer pathology. Bujingjiedu is an important lung cancer treatment principle. According to buingjiedu thought, Professor Zhang Yong Hong founded bujingjiedu recipe from his clinical experiences of many years. There were better clinical efficacy and high antitumor activity in the bujingjiedu recipe in early clinical and animal studies.[Objective] Based on previous studies, the changes of nuclear transcription factor NF-KB and AP-1 after the complements of the bujingjiedu treat on lung cancer A549 cells are researched.[Methods] Firstly, the monomer of the bujingjiedu recipe components (Cordyceps glycosides, Annona glycosides) is extracted. The inhibition of A549 cells is detected by MTT. The human lung cancer A549 cell apoptosis is observed by AlinexinV-FITC flow cytometry and transmission electron microscopy after treated with the components of bujingjiedu recipe.After the components of bujingjiedu recipe treat on the lung cancer A549 cell,the expression of NF-KB P65 is detected by Western Blot, the combination activity of NF-KB and AP-1 protein with DNA is discovered by Electrophoretic Mobility Shift Assay (EMSA).The luciferase values of NF-KB and AP-1 are detected with micro-plate luminescence after the components of bujingjiedu recipe treat on the lung cancer A549 cell transfected with plasmid contained pNF-KB-luc and pAP1-luc gene. The changes of NF-KB in people lung cancer A549 cell were observed with laser total focused microscope.[Results] Cordycepin and bullatacin were extracted from bujingjiedu. They can inhibit lung cancer A549 cells and the best time of cordycepin and bullatacin on lung cancer A549 cells was 72h in MTT. IC50 were 40.369 ug/ml and 30.893 ug/ml especially.After the components of bujingjiedu recipe treat on the lung cancer A549 cell the total apoptosis rate detected by AlinexinV-FITC flow cytometry were:Buiingjiedu 59.2%, Annona 53.2% and Cogrdyceps31.1%. Such as chromatin condensation, organelle degradation, mitochondrial swelling, cytoplasm swelling the typical apoptosis can be seen under the transmission electron microscope.Western results shows that the positive ratio of the positive gray-scale plot/internal reference values were:Bujingjiedu 0.862> Annona glycosides 0.656> Cordyceps glycosides 0.574.They are greater than the control group0.411.EMSA results showed that:the combination activity of NF-KB, AP-1 protein with DNA increased after treated with the components of bujingjiedu recipe. The positive gray scale plot of NF-KB in each group are Annona 233,571, Cordyceps 157,870, bujingjiedu 242437.5. They are higher than the control group 29897.5. The positive gray scale plot of AP-1 in each group are Annona 355,388,Cordyceps 244,311, bujingjiedu 318,776. They also are higher than the control group 84692.5.The results of luciferase values detected by micro-plate luminescence show that the NF-KB, AP-1 luciferase values increase significantly. There are significant difference compared with the control group (P<0.01).Observed under the laser confocal microscope the nuclear transcription factor NF-KB increased in the cytoplasm and the number of NF-K B into the nuclear increased mostly. It shows that the activity enhance.The order of the strength of the bujingjiedu effect on A549 lung cancer cells is bujingjiedu> bullatacin> cordycepin.[Conclusions] 1 The Annona glycosides and Cordyceps sinensis can induce lung cancer A549 cells apoptosis.2 The number of NF-KB and AP-1 increased and the activity enhanced after the components of bujingjiedu recipe treat on the lung cancer A549 cell. It may be one of the molecular mechanisms of lung cancer A549 cells apoptosis induced by the components of bujingjiedu recipe...