Study On The Mechanism Of Jiaweifei Compound Prescription In Reversing Epithelial-mesenchymal Transition Of Human Lung Adenocarcinoma A549 Cell

Objective:(1)To analyze Professor Pan Bo’s experience in treating lung cancer.(2)To study the effect of drug-containing serum of Jiaweifeifu decoction on proliferation,metastasis and invasion ability of human lung cancer A549 cells.(3)To explore the effects of drug-containing serum of Jiaweifeifu decoction on the expression of E-cadherin,Snail and Vimentin protein and m RNA,then prove the effect of Jiaweifeifu decoction on epithelial-mesenchymal transition.Methods:(1)To analyze Professor Pan Bo’s experience in treating lung cancer.(2)Human lung cancer A549 cells were cultured and divided into 7 groups: The experimental groups were treated with lung cancer A549 cells in the medium containing 10%,15% and 20% supplemented with the drug-containing serum of Jiaweifeifu decoction,and the control groups were treated with the culture medium containing 10%,15% and 20% drug-free rat serum,respectively.In blank group,lung cancer A549 cells were treated with fetal bovine serum medium containing 10%concentration.Results:(1)Professor Pan Bo believes that Qi and Yin deficiency and stasis and toxin internal node are the main pathogenesis of lung cancer,and the treatment method is to nourish qi and Yin,remove blood stasis and detoxify.According to this treatment method,Jiaweifeifu decoction is created as an empirical formula for lung cancer.The treatment of lung cancer should distinguish the pathogenesis,integrate the whole and treat by stages.(2)CCK-8 experiment showed that after treatment with A549 cells for 24 h,36h and 48 h,the survival rate of cells in each concentration group of supplemented Jiaweifeifu decoction medicated serum was significantly lower than that in control group and empty group(P<0.01).The results indicated that Jiaweifeifu decoction medicated serum had a significant inhibitory effect on the proliferation of lung cancer A549 cells.(3)The scratch test showed that compared with the same concentration control group and the blank group,the supplemented Jiaweifeifu decoction medicated serum had obvious inhibitory effect on the migration ability of lung cancer A549 cells(P<0.01),and the higher the concentration,the stronger the inhibitory effect.(4)Transwell cell metastasis and invasion experiments showed that the three experimental groups had significant inhibitory effects on the metastasis and invasion ability of lung cancer A549 cells(P<0.01).The inhibitory effect was strongest in 20% drug-containing serum group.(5)Western blot analysis showed that the expression of E-cadherin protein increased(P<0.05),Vimentin Snail protein decreased significantly(P<0.01).(6)In the three experimental groups,the expression of E-cadherin protein increased,and the expression of Vimentin and Snail protein decreased.(7)RT-PCR showed that m RNA expression of E-cadherin group increased(P<0.05),while m RNA expression of Vimentin and Snail decreased significantly(P<0.01).Conclusion:Professor Pan believes that the treatment of lung cancer should be holistic,integrated with traditional Chinese and western medicine,and phased.Jiawei Fei decoction medicated serum can inhibit the proliferation,invasion and migration of human lung cancer A549 cells.The drug-containing serum of Jiaweifeifu decoction may up-regulate the protein and m RNA expression of E-cadherin and down-regulate the protein and m RNA expression of Vimentin and Snail,thereby reversing the epithelial mesenchymal transformation process of human lung adenocarcinoma A549 cells.