Study of transfection of Livin antisense oligodeoxynucleotide on apoptosis of QBC939 cells and its mechanism

Abstract

Objective To study the effect of transfection of Livin antisense oligodeoxynucleotide (Livin ASODN) on Livin mRNA and Livin protein expression and apoptosis of QBC939 cells.Methods Livin ASODN were transfected into cell line QBC939 byLipofectamine TM 2000.60 h After transfection,to measure Livin mRNA expression by reverse transcriptase polymerase chain reaction (RT-PCR) and Livin protein expression by immunohistochemistry and confocal laser scanning microscopy ; the changes of cell apoptotic were detected by methyl thiazol tetrazolium (MTT).and the apoptosis of QBC939 cell was detected by Annexin V-PE flow cytometry (FCM) after the transfection.Results The mRNA and protein expression level of Livin in ASODN group was significantly lower than that in control group (P ＜0.05) (54.96 ± 3.00,88.31 ± 2.50,85.90 ± 0.62,90.36 ± 2.43) ; the results of MTT showed that the rate of cell apoptosis of QBC939 cells were the most obvious at 60 hours after Livin ASODN transfection (46.41 ± 1.13,3.10 ±0.44,2.66 ± 0.26,0) (P ＜ 0.05).the apoptosis of QBC939 cell was higher than those in control group after Livin ASODN transfection [(36.98 ±4.09)％,(3.77 ±0.25)％,(7.51 ±0.3)％,(6.83 ±0.23) ％].Conclusion The transfection of Livin ASODN inhibited Livin gene and protein expression,and Obviously induced apoptosis of Bile duct cancer QBC939 cells,which may be the new target for gene therapy in Bile duct cancer. Key words: Livin; Antisense oligodeoxynucleotide; Bile duct cancer; Cell apoptosis