Experimental Studies Of The Isomycin Derivant On Anti-proliferation And Inducing Apoptosis Of Human Lung Cancer A549 Cell Line

Objective To study the effects of the isomycin derivant on anti-proliferation and inducing apoptosis and its possible mechanism of human lung cancer A549 cell line to will provide the experimental basis for development of high performance anti-tumor medicine.Methods MTT assay was used to determine the effects of isomycin derivant on A549 human lung cancer cell growth inhibitory.Morphological changes of the apoptosis of A549 human lung cancer cells were observed by using inverted light,fluorescent microscope and transmission electronic microscope.Apoptotic rate was calculated with terminal deoxynucleotidy transferase-mediate dUTP nick-end labeling(TUNEL).The protein expressions of ki-67 and the apoptosis-related gene such as Bcl-2,Bax.Flow cytometry was used to detected cell cycle distribution of A549 cells and apoptotic rate. Results The MTT test showed that isomycin derivant in concentration of 5,10,20μg/ml could inhibit the hyperplasis of A549 human lung cancer cells in a doseage and time dependent manner.The A549 human lung cancer cells treated with isomycin derivant showed cellular volume small and round,nuclear chromatin condensation,loose between the cells and adherent ability decrease under invert microscope.The A549 human lung cancer cells in experimental group by HE staining showed morphological changes such as nuclear chromatin condensation,plasma menbrane bleb formation and the apoptotic body formation.Human A549 lung cancer cells stained by AO/BO double fluorescent staining were observed under fluorescent microscope,appearing as chromatin orange-red condensation in nuclear,orange-red or orange-yellow fluorescence disappeared and light green fluorescent apoptotic bodies around cells after induction of isomycin derivant,ki-67 anti-proliferation test indicated that the isomycin derivant of various concentration and time all inhibited A549 lung cancer cells in dose and time-dependent manner.The expression of bax was up regulated and Bcl-2protein was down regulated after exposure to isomycin derivant.Conclusions1) The human lung cancer A549 cells are inhibited significantly by the isomycin derivant in a dosage and time dependent manner.2) The isomycin derivant induces the apoptosis of human lung cancer A549 cells3) The isomycin derivant inhibits the growth of human lung cancer A549 cells by interfering the cell cycle,and the mechanism of molecular biology for inducing apoptosis of human lung cancer A549 cells is correlated with down regulation of apoptosis suppressor gene Bcl-2 and up regulation of promoting apoptosis gene Bax.