Circulation Eliminate Tumor Side Containing Serum On Human Lung Cancer A549 Cells, Fas / Fasl Expression

Objective: It has proved that the pharmaceutical serum containing Hua YuXiao Liu Fang has function of inducing the apoptosis of lung cancer cells. Toobserve the change of Fas and FasL in human lung carcinoma cell line(A549), to investigate the mechanism of the decoction of Hua Yu Xiao LiuFang in the induction of apoptosis of human lung carcinoma cell line (A549)by serologic pharmacological method and provide theory basis for thedecoction of Hua Yu Xiao Liu Fang to popularize and apply in clinic practice.Methods:(1)The rabbits were divided into five groups at random, There are fourrabbits in every group. Hua Yu Xiao Liu Fang were adminidtered po to rabbitsin different dose. Three days later, collecting blood through carotideatery, taking serum apart, freezing and storing in low temperature.According to the basic Traditional Chinese Medicine theory, the pathologiccharacteristic of lung carcinoma is stagnation of phlegm and turbid qi.(2)The human lung carcinoma A549 cell line was adopted as target cell.After incubating A549 cell and Chinese drug, mainly composed of nua Yu XiaoLiu Fang, borne rabbit serum at different dose with serologicpharmacological method.(3)Being treated with the five serums which mentioned above fou 24 and48 hours, the apoptosis morphological changes were observed under theinverted microscope. To get the number of Fas and FasL with means ofimmunohistochemical method and analyzed.Result:(1)With the inverted microscope, After being treated withdrugless-bearing serum. AS49 cell grew along and full of cell, grew freeless.After being treated with drug-bearing serum, A549 cell grew against thewall and then peeled off gradually, which showed the characters of cellsapoptosis like chromatin condensation. High dose serum of the decoctionof Hua Yu Xiao Liu Fang is the best. (2)With HE stain and light microscope, after being treated withdrug-bearing serum, A549 cell showed the typical apoptosis morphologicalchanges, such as shrinkage, crescent shape concentrated chromatin andfragmentation. High dose serum of the decoction of Hua Yu Xiao Liu Fangis the best. After being treated with drugless-bearing serum, A549 cellgrew very well, the typical apoptosis morphological changees could not beseen.(3)From the result of immunohistochemical method and analyzed, we can seethat the number of Fas has been raised after been treated with drug-bearingserum, while the number of FasL is in the opposite way. The number of Faswhich treated with high does drug-bearing serum at 24h or 48h wassignificantly higher than that of the control serum(P＜0.05).And the numberof Fas rised along the dose of the drug-bearing serum and delay of the effecttime. While The number of FasL which treated with high does drug-bearingserum at 24h or 48h was significantly lower than that of the controlserum(P＜0.05). And the number of FasL inhibited by the dose of thedrug-bearing serum and delay of the effect time.Conclusion: It demonstrates that the decoction of Hua Yu Xiao Liu Fang couldbe effective in lung carcinoma which was dependent on both time and dose.The above results suggest that the antineoplastic molecular mechanisms ofthe decoction of Hua Yu Xiao Liu Fang may be its effect on inhibiting Fasto restrain A549 cells proliferation and raise the number of FasL of A549cell. It provides theory basis for the decoction of Hua Yu Xiao Liu Fangto popularize and apply in clinic practice.