[The effect of the fused gene of SP-TAT-Apoptin transfected by lentivirus on HepG2 cells].
OBJECTIVE
To Package the recombinant lentivirus containing the fused gene of SP-TAT-Apoptin to infect HepG2 cell and measure the efficiency of apoptosis.


METHODS
The eukaryotic expression vector of SP-TAT-Apoptin fused gene and other packaging plasmids were transfected into 293FT cells by Lipofectamine(TM);2000 reagent. The supernatant of the cultured 293FT cells was harvested and virus titration was determined by real time PCR. The expression of the fused gene of SP-TAT-Apoptin in 293FT cells infected by the recombinant lentivirus was examined by immunofluorescence histochemistry method. At the same time, the apoptosis rates of the HepG2 cells infected by the recombinant lentivirus were determined by using flow cytometer.


RESULTS
The 293FT cells infected by the recombinant lentivirus could express the fused protein SP-TAT-Apoptin. Anexin-V PI assay showed that SP-TAT-Apoptin carried by the recombinant lentivirus could cause the HepG2 cell apoptosis, and its apoptosis rate was significantly more than paired control group and SP-TAT-Apoptin carried by liposomes only.


CONCLUSIONS
The recombinant lentivirus of SP-TAT-Apoptin is successfully packaged and it can induce HepG2 cells to apoptosis.
Han
Su-xia
SX
Oncology Center of The First Affiliated Hospital, College of Medicine, Xi'an Jiaotong University, Xi'an 710061, China. huansuxia22099@yahoo.com.cn
Zhao
Jing
J
Ma
Jin-lu
JL
Huang
Chen
C
Lü
Yi
Y
Ou
Wei
W
Jia
Xi
X
D017209:Apoptosis / Q000235:genetics; D019677:Artificial Gene Fusion; D036022:Capsid Proteins / Q000235:genetics*; D004274:DNA, Recombinant / Q000235:genetics; D005434:Flow Cytometry; D015696:Gene Products, tat / Q000235:genetics*; D015316:Genetic Therapy; D056945:Hep G2 Cells; D006801:Humans; D016086:Lentivirus / Q000235:genetics* / Q000502:physiology; D009369:Neoplasms / Q000235:genetics / Q000628:therapy / Q000821:virology; D021382:Protein Sorting Signals / Q000235:genetics*; D014162:Transfection / Q000379:methods*
D036022:Capsid Proteins; D004274:DNA, Recombinant; D015696:Gene Products, tat; D021382:Protein Sorting Signals; C091717:VP3 protein, Chicken anemia virus