Inhibition of mouse hepatocyte apoptosis via anti-Fas ribozyme.
OBJECTIVE
To investigate the effects of anti-Fas ribozyme on Fas expression and apoptosis in primary cultured mouse hepatocytes.


METHODS
Mouse hepatocytes were isolated by using collagenase irrigation. A hammerhead ribozyme targeting the Fas mRNA was constructed, and transfected into mouse hepatocytes via Effectene. Then Fas expression in mouse hepatocytes was detected by RT-PCR and western blotting. After being treated with anti-Fas antibody (JO2), hepatocytes viability was measured with MTT assay. Caspase-3 proteolytic activity was detected, and cell apoptosis was measured according to Annexin V-FITC apoptosis detection kit.


RESULTS
Fas expressed in primary mouse hepatocytes. Fas expression in hepatocytes transfected with anti-Fas ribozyme was decreased remarkably and correlated with the resistance to Fas-mediated apoptosis as determined by flow cytometry and caspase-3 proteolytic activity.


CONCLUSIONS
Anti-Fas ribozyme can remarkably decrease the Fas expression in mouse hepatocytes, thus inhibit Fas-mediated apoptosis in hepatocytes. It is suggested that anti-Fas ribozyme could significantly increase the resistance of transplanted hepatocytes to apoptosis and improve the survival of transplanted hepatocytes.
Zhang
Min
M
Institute of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, Hubei Province, China.
He
Wei
W
Liu
Fang
F
Zou
Ping
P
Xiao
Juan
J
Zhong
Zhao-Dong
ZD
Hu
Zhong-Bo
ZB
D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D002470:Cell Survival / Q000502:physiology; D002478:Cells, Cultured; D015870:Gene Expression; D022781:Hepatocytes / Q000166:cytology*; D051379:Mice; D016337:RNA, Catalytic / Q000235:genetics*; D014162:Transfection; D019014:fas Receptor / Q000235:genetics*
D016337:RNA, Catalytic; D019014:fas Receptor; C427204:hammerhead ribozyme; C505409:Casp3 protein, mouse; D053148:Caspase 3; D020169:Caspases