Effects Of Livin Antisense Oligonucleotide On The Proliferation, Apoptosis And Drug Resistance Of K562Cells

Objective:To study the effects of Livin antisense oligonucleotide (ASODN) on the proliferation and apoptosis of human leukemia(K562) cells and investigate whether antisense oligonucleotide can sensitize leukemia cells to chemotherapy. It maybe provide experimental evidence to Livin gene targeted for the clinical treatment of leukemia.Methods:Specific phosphorothioate ASODN and MSODN target Livin mRNA were synthesized and transfected into K562cells following cationic liposome. To study the effects of Livin ASODN on the proliferation of K562cells, cells were divided into4groups:Lip-ASODN group, Lip-MSODN group, Lip control group and cell control group. The proliferation inhibition of K562cells was assessed by MTT after24and48hours. To study the effects of Livin ASODN on the apoptosis of K562cells, cells were divided into4groups: Lip-ASODN group, Lip-MSODN group, Lip control group and cell control group. The apoptosis rate of each group was detected by Annexin V-FITC after24and48hours. To study the effects of Livin ASODN on the expression of Livin mRNA in K562cells, cells were divided into4groups:Lip-ASODN group, Lip-MSODN group, Lip control group and cell control group.The expression of Livin mRNA was detected by reverse transcription polymerase chain reaction (RT-PCR) after24hours. To study the effects of Livin ASODN on the drug resistance of K562cells, cells were divided into3groups:HHT-ASODN group, HHT-Lip group, HHT-cell control group. The IC50of K562cells to HHT was assessed by MTT after48hours. To study the effects of Livin ASODN combined HHT and low-dose Ara-C on the proliferation of K562cells, cells were divided into4groups:HHT group, Ara-C group, HHT-Ara-C group, HHT-Ara-C-ASODN group. The Cell survival rate of K562cells was assessed by MTT after12hours,24hours and48hours.Results:The proliferation of K562cells was inhibited significantly by Livin ASODN in a time-dose-dependent manner. Livin ASODN at a final concentration of600nmol/L could inhibit the K562cells proliferation after48hours, IR=(52.99±2.67)%. It was significantly higher contrast to other three groups (P<0.01). Livin ASODN at a final concentration of600nmol/L could inhibit the expressions of Livin mRNA and the apoptosis rate reached (36.89±1.08)%(P<0.01),but the differences between Lip-MSODN group,Lip control group and cell control group were not statistically significant(P>0.05).Livin ASODN could enhance sensitivity to HHT in K562cells, IC50was (0.37±0.02) mg/L. Livin ASODN combined HHT and low-dose Ara-C could inhibit the proliferation of K562cells apparently (P<0.01).Conclusions:Livin ASODN inhibit Livin gene expression and the proliferation of K562cells effectively, enhances the apoptosis and sensitivity to HHT in K562cells.Livin gene will maybe an appropriate and effective target for the gene therapy of leukemia.