Effects Of The Biological Characters By Using Salinomycin With A549Lung Adenocarcinoma Cell Lines

Objective:Salinomycin is a polyether antibiotic,which is a member of ionospherethat isolated from streptomycin albus. Recently,it has been shown to kill breast cancerstem cell and inhibit human breast cancer cell growth and metastasis specificity. Cancerstem cells (CSC) theory is that cancer stem cells is a small fraction of cancer cells whichhave multilineage differentiation potential,unlimited proliferation ability, self-renewalability.At the same time,CSCs plays an important role in the development of tumor. Theside population cells(SP) contain most part of cancer stem cells.Stem cell transcriptionfactor Oct4is one of the POU transcription factor family members that is a cellpluripotent differentiation marker. Oct4has the ability to regulate the progress of stemcells.At the present,the mechanism of salinomycin killing CSCs is remain unclear.Sowe detected the effect of salinomycin on lung adenocarcinoma cell lineA549,furthermore,we discuss the mechanism of salinomycin’s function to A549.Methods: Serum-free culture of human lung adenocarcinoma cell line A549cellstumor cells ball. With different concentrations of salinomycin role in cultured A549cells of lung adenocarcinoma and lung adenocarcinoma A549cells ball. The scratch testdetection of different concentrations of salinomycin on lung adenocarcinoma A549cellsin vitro migration ability; using Transwell assay for the detection of differentconcentrations of salinomycin on lung adenocarcinoma A549cells invasion ability;using CCK8assay for the detection of different concentrations of salinomycin on lungadenocarcinoma A549cells survival rate; Flow cytometry using different salinomycinconcentration on lung adenocarcinoma cell ball side population proportion of influence.Western-blot method for the detection of different concentrations of salinomycin on lung adenocarcinoma A549cells the expression of Oct4protein.Results:1Serum free culture medium with epidermal growth factor (EGF), basicfibroblast growth factor (bFGF) were cultured, after14days of culture developed lungcancer stem cell balls.2In10hours after administration of the following results: negative control groupmobility of distance70um,30ug/ml salinomycin treatment group40um,60ug/mlsalinomycin treatment group10um. Tips of salinomycin can obviously inhibit lungadenocarcinoma A549cells migration. And as the drug concentration increased,inhibition was enhanced.3In12hours after administration of the following results; the negative controlgroup, the onset of invasive cells the number of (32.3+2.5)/HP,30ug/mlsalinomycin treatment group (21.4+1.8)/HP,60ug/ml salinomycin treatment group(1.3+0.3)/HP. Tips of salinomycin can obviously inhibit lung adenocarcinoma A549cells invasion.4Results show that, with the increasing concentration of salinomycin medication,lung adenocarcinoma cell survival ratio was significantly decreased, that salinomycinon lung adenocarcinoma A549cells5Salinomycin treatment on detection results: joining the60ug/ml measured sidepopulation proportion is3.4%, but join30ug/ml groups measured side populationproportion is10.5%. A Not to join the Salinomycin side population proportion is12.5%,B Not to join the Salinomycin (join the blocker reserpine), side populationproportion is0.7%. in adding salinomycin groups, Can be found, along with theincrease of concentration, the ratio of side population decline trend.6Western Blot results show, with salinomycin in lung adenocarcinoma celldosage increases, the amount of Oct4expression is gradual decline. In clinicalspecimens, adenocarcinoma of the lung tissue Oct4expression, no expression in normallung tissue.Conclusion:1With the increase of concentration of salinomycin can affect lungadenocarcinoma A549cell invasion, migration ability and reduce lung cancer survivalrates.2With the increase of concentration of salinomycin to Salinomycin can reducepulmonary adenocarcinoma A549cell ball side population cell ratio. 3Salinomycin to with the increasing concentration of reduced stem celltranscription factor Oct4in lung adenocarcinoma by expression.