Study Of Transfection Of Livin Antisense Oligodeoxynucleotide On The Proliferation Of Bile Duct Cancer QBC939 Cells

Objective:to study the effect of transfection of livin antisense oligodeoxynucleotide (Livin ASODN) on Livin mRNA and Livin protein expression of QBC939 cell; to study the effect of transfection of livin antisense oligodeoxynucleotide(Livin ASODN) on the proliferation and apoptosis of QBC939 cells.Methods:The 5'end of Livin ASODN was labeled with FITC,then Livin ASODN were transfected into cell line QBC939 by LipofectamineTM 2000,to observe the cell of Livin ASODN transfection by Fluorescence microscope, and calculate the rate of transfection; The experiment was divided into 4 groups:Livin ASODN group (Antisense Oligodeoxynucleotide group),Livin NSODN group (Nonsense Oligodeoxynucleotide),Lipofectamine group,Control group.Livin ASODN/NSODN were transfected into QBC939 cell line by LipofectamineTM 2000. the changes of cell proliferation were detected by MTT,and select the best rection time and concentration;to measure Livin mRNA expression by RT-PCR after the transfection; to measure Livin protein expression by immunohistochemistry and confocal laser scanning microscopy after the transfection; and the apoptosis of QBC939 cell was detected by Flow cytometry (FCM) after the transfection.Result:Under the help of LipofectamineTM 2000,Livin ASODN can be successfully transfected into QBC 939 cell line.and transfection efficiency is high, which increased as the concentration was elevated within a certain range.the highest efficiency was at 24 hours after 500nmol/L Livin ASODN transfection(70.50±2.08). the results of MTT showed that the inhibition of proliferation of QBC939 cells was increased as the concentration and infection time was elevated with a certain range(50-500nmol/L), and the most obvious(46.41±1.13)was at 60 hours after 500nmol/L Livin ASODN transfection (p< 0.05).the level of Livin mRNA and Livin protein expression in ASODN group was obviously lower than that in control group at 60 hours after livin ASODN transfection(p< 0.05); the apoptosis of QBC939 cell was higher than those in control group at 60hours after Livin ASODN transfection.Conlusion:the transfection of Livin ASODN inhibited Livin gene expression, and obviously inhibit the proliferation of QBC939 cell and induced apoptosis of QBC939 cells, which may be the new target for gene therapy in Bile duct cancer.Experiment show that the critical region of gene may be blocked by Livin ASODN and can inhibit the proliferation of QBC939 cell and to induce it apoptosis.