Inhibition of survivin expression to induce the apoptosis of hepatocarcinoma cells by adenovirus-mediated siRNA.
In order to provide more efficient transduction of plasmid siRNA into target cells, develop more susceptible transduction into cancer cell types, and more easily explore application in animal experiments, we examined development of an adenoviral vector-mediated siRNA expression system and inhibition of survivin gene expression to induce the growth and apoptosis of hepatocarcinoma cells. A system of adenoviral vector-mediated siRNA expression was constructed for the survivin gene. Survivin gene expression in HepG2 cells infected with recombinant adenovirus was detected by Western blot and RT-PCR, and apoptotic cells were investigated by FAC. Western blot analysis showed that the infection of adenovirus-mediated siRNA against survivin efficiently inhibited the expression of survivin in hepatocarcinoma cells with an inhibitory rate of 66.32%. Semi-quantitative RT-PCR showed that survivin gene mRNA transcription was reduced by nearly 72.34% with a peak at 72 h. The number of apoptotic cells increased. In conclusion, results demonstrated that this adenovirus-mediated siRNA system could serve as a useful tool for both basic research on the analysis of gene function and cancer therapy applications.
Yan
Ge
G
Shandong Institute of Parasitical Diseases, Shandong, China. gey919@yahoo.com.cn
Duan
Ruihong
R
Yin
Kun
K
Zhu
Song
S
Liu
Qiaoqiao
Q
Gong
Maoqing
M
Wang
Huaiwei
H
Sun
Chuanhong
C
Pu
Dan
D
Tang
Ni
N
Huang
Ai-Long
AL
D018248:Adenoma, Liver Cell / Q000378:metabolism*; D017209:Apoptosis / Q000235:genetics / Q000502:physiology; D015153:Blotting, Western; D045744:Cell Line, Tumor; D005434:Flow Cytometry; D006801:Humans; D020287:In Situ Nick-End Labeling; D008113:Liver Neoplasms / Q000378:metabolism*; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D034741:RNA, Small Interfering / Q000235:genetics / Q000502:physiology; D020133:Reverse Transcriptase Polymerase Chain Reaction
D008869:Microtubule-Associated Proteins; D034741:RNA, Small Interfering