The Experimental Research Of Parthenolide On Changing The Cell Cycle And Influencing The Apoptosis Pathways Of Human Lung Cancer A-549Cell Line

Objective:To investigate the effect of parthenolide on inhibiting proliferation and inducing apoptosis of human lung cancer A-549cell line in vitro.and to provide the evidence of experiment and theory for the new anti-tumor Chinese medicine.Methods; It was applied that training human lung cancer A-549cells in vitro. MTT assay was used to measure the toxic effect and the growth inhibition rate of human lung cancer A-549cells which were treated with parthenolide by different concentrations; Morphological changes of the tumor cells were observed by inverted microscope, HE staining, AO/EB double staining and Annexin V-FITC/PI double staining; The method was used that testing the level of reactive oxygen species in cells and discussing the relationship between ROS with the apoptosis of cells;Flow cytometry was used to investigate the cell cycle distribution and induce the apoptosis of situation; Immunocytochemistry was used to detect the changes of protein expression and in order to further explore the mechanism research of parthenlide on inducing the proliferation and apoptosis of human lung cancer A-549cell line.Results:(1) MTT assay showed that parthenolide by different concentrations had different levels of growth inhibition on human lung cancer A-549cell line and the effect was in a doseage and time gradient dependence.(2)Under inverted microscope observed that compared with the control group, A-549cell showed decreased and gone round.chromatin gathered,also the connection between the cells loosed and easied to fall off; HE staining showed that the nucleus of A-549cell with parthenolide exhibited pyknosis,part of the cell showed apoptosis; The AO/EB double staining and microscope demonstrated that the chromatin of living cells were green but along with the extension of time that were treated with parthenolide, the early apoptosis cells were yellow green and the late apoptosis and the dead ones were orange; Annexin V-FITC/PI double staining and fluorescence microscope showed the control group cells almost colored,but the experimental group cells appeared amount of green and red fluorescent apoptosic cells.(3)Reactive oxygen species assay detected the level of the ROS in cells increased significantly that compared with the control group.(4)Flow cytometry showed that the percentage of cells in GO/G1phase decreased and the percentage of cells in S&G2/M phase increased after treated with parthenolide,it pointed out parthenolide could arrest A-549cells in S&G2/M phase;(5)Parthenolide up-regulated the expression of Caspase-3、Caspase-8、 Caspase-9、 Fas、 Bax, it also dowm-regulated the expression of Bcl-2、Ki-67、 Livin、 Survivinand NF-κB in experimental groups, and they were markedly different from the control groups(P<0.05).Conclusions:1. Parthenolide has obvious inhibited the proliferation of A-549cells in vitro,and the effect is a concentration and time dependence,it arrest the A-549cells in the S&G2/M phase.2. The A-549cells after were roled with parthenolide show the obvious changes of apoptosis,the mechanism of apoptosis which parthenolide induce is completed by the death receptor pathways and the mitochondrial pathways,NF-KB may participate the signal transduction that parthenolide induced the apoptosis of A-549cells.3. The increase of the level of the ROS that were treated with parthenolide may participate in inducing the apoptosis mechanism.4. Parthenolide has the antineoplastic effect, we hope it will be exploded and be employed as a new anti-tumor medicine in the future.