Effects Of Diao Liu Pillon Apoptosis Related Gene Caspase-3 Expression Of MCF-7human Breast Carcionma Cell

Object: Study the effects of induced apoptosis of DiaoLiu Pill on MCF-7 in vitro and investigate the mechanism.Provide the basis and the train of thought for the future study of Chinese medicine anti-cancer mechanism,provide certain basis of molecular biology for the clinical treatment of breast cancer and offer the corresponding theoretical basis for the application of this drug in clinical.Methods: Human breast cancer cell lines MCF-7 are Cultivated with MEM cultures contained by 1% of penicillin,streptomycin,1% fetal bovine serum at 37?and 5%CO2 incubator.1.The method of morphological observation with light microscope :Take out the cell sheets after culturing the cell for 48 hours with DiaoLiu Pill extract,Proceed with HE staining,then observe them with light microscope and record them.2.MTT-test: Cell lines MCF-7 were vaccinated in 96-well plates at 1×105 cells per well,setting up five complex holes and randomly divided into six groups:,DiaoLiu Pill of high concentration group,DiaoLiu Pill of medium concentration group,DiaoLiu Pill of low concentration group,negation control group,FAM positive control group.Carry out MTT-test,calculate the proliferation inhibition rate and select the most suitable drug concentration.3.Apoptosis of cell lines MCF-7 after given drug were observed by Hoechst33258 staining under fluoroscope microscope.4.Cultivate all of the groups for 48 h,collect the cell after digested by trypsin,then extract their RNA.According to the quantity of RNA,test the quantity of caspase-3 in the different groups.Results: DiaoLiu Pill can inhibit the proliferation of cell lines MCF-7 and the inhibition depends on the dose.The inhibition rate of tumor of the high concentration group approximates the TAM positive control group.Compared with negation control group,the inhibition rate of DiaoLiu Pill group,especially DiaoLiu Pill of high concentration group,was significantly higher(P<0.05);DiaoLiu Pill groups are more apoptotic bodies than negation control group underfluoroscope microscope.According to the test of RT-PCR,DiaoLiu Pill groups can increase the quantity of caspase-3and the increment depend on the dose.Compared negation control group with DiaoLiu Pill groups,there are remarkable statistical difference(p<0.05).Conclusion: DiaoLiu Pill can inhibit the proliferation and promote apoptosis of cell lines MCF-7.The mechanism may be by increasing the lever of caspase-3 resulting in suppression and apoptosis of breast cancer cell.