The Effects On Compound Components Of Shanchanteng On Proliferation And Apoptosis Of Lung Cancer And Its Mechanism

Research purpose:To research the effects of compound components of Shanchanteng on the proliferation and apoptosis of Lung cancer cells, the regulation of apoptosis related protein and effects on Lung cancer cell morphology, to investigate the effective molecular mechanism of compound components of Shanchanteng against lung cancer.Research methods:1.To detected in inhibiting the proliferation of Taxus monomer, toad skin monomers, monomers of Tripterygium wilfordii on A549cells and NCI-H1688cells by using MTT method, to compare their anti-cancer activity.2.Apoptosis was detected by flow cytometry, observed the influence of Taxus monomer, toad skin monomers, monomers of Tripterygium wilfordii on A549cells and NCI-H1688cells in cell cycle.3.To detect the effect of immunohistochemistry fir toad rattan compound components of lung cancer associated protein Bad, Bax, Bcl-2protein regulation.4.Transmission electron microscopy observation on A549cells in vitro and NCI-H1688cell morphology and observe whether apoptotic bodies, nuclear chromatin pyknosis, margination phenomenon.5.Laser confocal microscopy was used to observe the changes of compound components of Shanchanteng composition effects on A549cells and NCI-H1688cells.Results:1.Cell drug24, after48hours, compared with the blank control group, different concentration and duration of the three monomers on A549cells and NCI-H1688cells were inhibited (P<0.05), especially the inhibitory effect of high dose group significantly (P<0.01),and in the same concentration, with the extension of time, three inhibited the growth of A549cells was monomer and NCI-H1688cells increase.2.A549cells and NCI-H1688cells after different doses of three monomer48h after treatment, each dose group showed varying degrees of apoptosis, and the percentage of apoptotic cells in the early, middle and late show with the dose increased and growth. The total cell apoptosis rate in each dose group compared with the control group, the difference was statistically significant (P<0.05).3.Immunohistochemistry showed that the anti-apoptotic Bcl-2protein expression decreased, increased total Bax protein/Bcl-2protein, apoptosis Bad protein; apoptosis suppressing gene Bcl-2downregulation of mRNA expression, increase in total Bax mRNA/Bcl-2mRNA Bad mRNA expression, apoptosis.4.Transmission electron microscope observation showed:three monomers of A549cells and NCI-H1688cells after48h in size Shrinkage, the membrane is not complete, the nuclear membrane disappeared, nuclear atrophy, fragmentation, individual and formation of apoptotic bodies; part of chromatin pyknosis, along the nuclear membrane forms a plurality of crumb edge set; cell swelling, vacuole increased significantly. The cell membrane in the normal rats, complete with smooth surface, microvilli, cytoplasmic density high, no bubble, nuclear chromatin distribution, large nucleolus more complete, center or edge shift; mitochondrial ridge broken or disappeared.5.Laser confocal microscopy to compound components of Shanchanteng with A549cells and NCI-H1688cells, there are varying degrees of phosphatidylserine eversion, apoptosis phenomenon.Conclusions:1. Compound components of Shanchanteng (Taxus monomer, toad skin monomers, monomers of Tripterygium wilfordii) in vitro on lung adenocarcinoma A549cells and small cell lung cancer NCI-H1688cells can inhibit proliferation and suppress the dependence rate, concentration and time of.2.Compound components of Shanchanteng can induce A549cell and NCI-H1688cell apoptosis, and induce cell has the corresponding morphological changes.3.The mechanism of A549cell and NCI-H1688cell apoptosis induced by Sugi To compound components of regulatory proteins may also be associated with regulation of Bcl-2family Bax, Bcl-2, Bad protein and Bax of mRNA, Bcl-2mRNA, Bad mRNA expression.