Survivin antisense oligonucleotide induces human Hep-2 cell apoptosis
JI Yu-xi
Abstract
JI Yu-xi
Abstract
Objective Survivin highly overexpresses in the most of human tumors, and it may play an important role in the development of tumor. The aim of this study was to explore the effects of survivin antisense oligonucleotide(ASODN)on the proliferation and the apoptosis of human Hep-2 cell. Methods Hep-2 cells were transfected with survivin ASODN mediated by lipofectamine, MTT [3-(4,5-dimethyldiazol-2-yl)-2, 5 dipheny tetrazolium bromid]method was used to observe the cell growth inhibitory rate, the expressions of survivin mRNA and protein were detected by reverse transcriptase polymerase chain reaction(RT-PCR) and Western blot respectively. Flow cytometry was used to examine cell apoptosis rate. Kinase activity test was used to detect the changes of caspase-3 activity. Results Survivin ASODN obviously inhibited the cell growth of Hep-2 cells after transfection. After transfected with survivin ASODN the expressions of survivin mRNA and protein of Hep-2 cells were down-regulated, and apoptosis rate was significantly increased. The activity of caspase-3 increased highly in Hep-2 cells transfected with survivin ASODN, which showed time-dependent. Conclusions Survivin ASODN could inhibit the proliferation of Hep-2 cell and induced apoptosis through down-regulating the the expressions of survivin mRNA and protein.
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Objective Survivin highly overexpresses in the most of human tumors, and it may play an important role in the development of tumor. The aim of this study was to explore the effects of survivin antisense oligonucleotide(ASODN)on the proliferation and the apoptosis of human Hep-2 cell. Methods Hep-2 cells were transfected with survivin ASODN mediated by lipofectamine, MTT [3-(4,5-dimethyldiazol-2-yl)-2, 5 dipheny tetrazolium bromid]method was used to observe the cell growth inhibitory rate, the expressions of survivin mRNA and protein were detected by reverse transcriptase polymerase chain reaction(RT-PCR) and Western blot respectively. Flow cytometry was used to examine cell apoptosis rate. Kinase activity test was used to detect the changes of caspase-3 activity. Results Survivin ASODN obviously inhibited the cell growth of Hep-2 cells after transfection. After transfected with survivin ASODN the expressions of survivin mRNA and protein of Hep-2 cells were down-regulated, and apoptosis rate was significantly increased. The activity of caspase-3 increased highly in Hep-2 cells transfected with survivin ASODN, which showed time-dependent. Conclusions Survivin ASODN could inhibit the proliferation of Hep-2 cell and induced apoptosis through down-regulating the the expressions of survivin mRNA and protein.
Key concepts: Survivin, Transfection, Lipofectamine, Apoptosis, Molecular biology, Cell growth, Chemistry, Cell