[SP-TAT-Apoptin induces G1 arrest in HepG2 cells].
OBJECTIVE
To investigate the effect of SP-TAT-Apoptin in inducing HepG2 cells apoptosis and the possible application on hepatocellular carcinoma gene therapy.


METHODS
Recombinant gene SP-TAT-Apoptin was amplified by PCR and cloned into the eukaryotic vector plenti6-V5-D-TOPO. After the recombinant plasmid was identified by restriction enzyme digestion analysis and DNA sequencing, CHO cells were stably transfected with SP-TAT-Apoptin gene and the culture supernatant was collected. Then the expression of the fusion protein was detected by RT-PCR and Western blot. HepG2 cells were co-cultured with the supernatant. At various times post co-culture, HepG2 cells were detected by FCM.


RESULTS
The secretory Tat-Apoptin has an additive bystander effect as an anti-cancer therapy in vitro. The recombinant Apoptin was able to be secreted from transfected cells and re-enter adjacent un-transfected HepG2 cells, it can induce HepG2 cells apoptosis and induce G0/G1 arrest.


CONCLUSIONS
SP-TAT-Apoptin can induce HepG2 cell apoptosis and cell cycle G1 arrest.
Han
Su-xia
SX
The First Affiliated Hospital Oncology Center, Xi'an 710061, China.
Ma
Jin-lu
JL
Lu
Yi
Y
Huang
Chen
C
Duan
Kang-min
KM
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D016466:CHO Cells; D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D002453:Cell Cycle / Q000187:drug effects*; D002477:Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D006224:Cricetinae; D003412:Cricetulus; D056945:Hep G2 Cells; D006801:Humans; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D054322:tat Gene Products, Human Immunodeficiency Virus / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*
D036022:Capsid Proteins; D011993:Recombinant Fusion Proteins; C091717:VP3 protein, Chicken anemia virus; D054322:tat Gene Products, Human Immunodeficiency Virus