The Study On The Cytotoxic Effect Of CIK Cells Combined With Icotinib Against Lung Adenocarcinoma Cells

BackgroundAccording to statics,lung cancer occupies the first place in the maligant tumor in China’scause of death,serious impact on people’s lives and health.Among them,non small cell lungcancer according to is the most common type of lung cancer,accounting for about80%~85%.Due to the lack of effective early examination and screening for lung cancerpatients,resulting in80%of our already advanced in the discovery and diagnosis,missed thebest treatment time and treatment methods.Operation is the first choice for cancer patients;tohave do not have operation of the syndrome of patients,radiotherapy and chemotherapy,isalso a very important method of treatment.But in recent years,due to the incompleteness ofthe operation,low sensitivity to radiotherapy and chemotherapy multidrug resistance and theclinical effect of the treatment,the poor.Actively explore people,emerge an the times requiretumor biotherapy,become the fourth treatment mode of comperhensive therapy oftumor.Among them,cytokine-induced killer cells(CIK) is the main effect of adoptiveimmunotherapy of tumor cells,become a research hotspot.CIK cells by mobilizing andstrengthening the immune function of the patients,so as to kill tumor cells and preventmetastasis and recurrence of the objective,improve the prognosis of cancer patients,thepatients’ quality of life have greatly improved.In addition,in recent years the cell receptor andmolecular regulation, the key base for targeting therapy has been studied and developed very well.In lung cancer,most people study is EGFR(epidermal growth factor receptor-tyrosinekinase inhibitor),such as erlotinib,gefitinib than,in clinical on achieved fairly good effect. Atthis stage,tumor treatment that an effective integrated treatment mode.Through thisexperiment,CIK cells and domestic EGFR-TKI(Icotinib) joint action, in the in vitro killingeffects on lung adenocarcinoma cell line, the feasibility study of two methods andefficiency,to provide guidance for clinical treatment of lung cancer in general.ObjectiveWe research and observate that CIK cell therapy and different concentrations of icotinibhydrochloride(Icotinib) on lung adenocarinoma cell lines:PC9cells(epidermal growth factorreceptor of mutant cells) and A549cells(epidermal growth factor receptor of wild-type cells)in vitro cytotoxic effect.MethodsIn cultured PC9cells and A549cells as target cells, the experiment was divided into fourgroups:CIK group, three different concentrations of icotinib hydrochloride group, CIK cellcombined with Icotinib group and blank control group (i.e. the target cells alone group).(1) CCK8kit for detection of CIK cells and Icotinib respectively and the combination onPC9cell and A549cell growth inhibition in vitro;(2) Annexin V-FITC kit using flow cytonetry to detect CIK cells and the influence of Icotinibon two kinds of target cell apoptosis;(3) The IC50were detected by MTT method of icotinib on two kinds of targeted cells.Results1CIK cells to two targeted cells killing effect comparison of CCK8method and PC9cell andA549cell killing rate and apoptotic rate of a floe cytometry, CIK had no significantdifference between two kinds of cytotoxic effect on targeted cells(P>0.05), that killer CIK cells on lung cancer cells is effective.2Comparison of CCK8on CIK cells combined with Icotinib group and the individual groupkiller and Annexin V-FITC apoptosis could be observed: the role of CIK cell+icotinibcombined group on targeted cell killing in vitro were significantly higher than that in thetreated group, the difference was ststistically significant(P<0.05).3Comparative statistics from experimental data analysis between PC9cells and A549cells:the combined group and Icotinib alone on PC9cells killing rate and apoptosis rate werehigher in the A549group,combined group on PC9cell killing rate reached65.85±1.44%,A549cells combined with killing rate reached52.92±1.57%, the differencewas statistically significant(P<0.05).4Two kinds of cells of the IC50value of icotinib on half of the PC9and A549cells after48hinhibition concentration(IC50) were0.21±0.03μ mol/L、4.79±0.74μ mol/L， thedifference was statistically significant(P<0.05), that targeted drugs on tumor cell effect ocEGFR gene mutation is better.ConclusionEach group after the cprresponding processing the data changes can be seen:CIK cell killingeffect on lung cancer cell can be applied in clinical lung cancer.EGFR gene targeted CIKcells combined with domestic drug in vitro can obviously inhibit the growth of lung cancercells,the effect of the combined therapy alone is better than traditional therapy,except tumorfor clinical outside new feasibility and selection.Cellular immune therapy and moleculartargeted drug combination,improve the treatment effect,enhance the patient’s quality oflife,reduce chemotherapy side, will be the new trend of the future of comprehensive cancertreatment.