Apoptosis Of Human Lung Adenocarinoma Cell Line A549Induced By Trametes Robiniophila

Objective:We chose human lung adenocarcinoma A549as research object, studied the effect of Trametes Robiniophila（TR） on growth inhibition and apoptosis induction of cell line A549using different methods, and pilot-studied the mechanism of apoptosis induction by TR,which will probide a basis for further investigating the molecular mechanism of lung cell apoptosis induced by TR, and will provide the theoretical and experiments evidence to treat lung cancer by TR.Methods:The inhibitory effect of Trametes Robiniophila on growth of A549cell line was examined by methyl thiazolyl tetrazolium （MTT） assay. Cell morphology was observed under reversed phase contrast microscope and reversed fluorescence microscope. Effect of TR on apoptosis induction of A549was analyzed by agarose electrophoresis.Flow cytometry showed cell cycle and apoptotic cell.The p53and bcl-2family expression in A549cells induced by Trametes Robiniophilaby by Western blot.Results:Results abtained by MTT indicated that inhibitory effect of the extracts on A549cells increased with the increase of concentration and time.The effect reached a peak when the concentration is3mg/ml.The inhibitory ratio of cell growth as the concentration of3mg/ml was39.3%,which showed negligible difference（P>0.05） compared with cisplatin.Change of cell morphology was obvserved under reversed phase contrast microscope and fluorescence microscope.The cell membrane looked whole but had blisters is the surranding areas.It looked shriveled and sunken, and apoptotic corpasculum appeared at the late stage;Fluorescent staining showed cell heavily-dyed dense fluorescent particles,reascent change, pyknotic or fragmented cell nuclears.The results of DNA electrophoresis analysis showed typical DNA ladders.The results of flow cytomotry showed that typical sub-diploid apoptosis peak was demonstrated treated with TR. A549cells were accumulated at G₀/G₁period while S period ratio fell Analysis results of Western blot:The expression level of p53protein in cell line A549treated by TR was increased while that of Bcl-2protein was decreased.