[Study on the in vitro cleavage abilities of ribozymes specific to different sites of bcr-abl fusion gene and their induction of apoptosis in K562 cells].
OBJECTIVE
To investigate the in vitro cleavage abilities of ribozymes specific to different sites of bcr-abl fusion gene and their effects on K562 cell.


METHODS
First, three single-ribozymes specific to the fusion point were designed. After recombination, 6 vectors including single-, double- and triple- ribozymes were constructed and their in vitro cleavage abilities were compared. Then the triple-unit ribozyme retroviral vector was transfected into K562 cell to test its effect on cell cycle, apoptosis and cell structure.


RESULTS
These ribozymes can cleave the template in vitro with different efficiency. The triple-unit ribozyme, with an efficiency of 70.8%, was the most efficient one. The cleavage efficiency of single-unit RZ1, RZ2 and RZ3 was 54.6%, 25.3% and 3.6%, respectively. Those of double-unit RZ12 and RZ23 were 60.7% and 30.3% respectively. The triple-unit ribozyme could inhibit the K562 cell growth by inducing apoptosis.


CONCLUSIONS
It is a new way to treat CML by ribozymes specific to bcr-abl fusion gene, which made it available to purge bone marrow by bcr-abl specific ribozymes.
Feng
Q
Q
Department of Hematology, Xijing Hospital, The Fourth Military Medical University, Xi'an 710033, China.
Sun
B
B
Zhao
Y
Y
D017209:Apoptosis; D001665:Binding Sites; D002453:Cell Cycle; D016044:Fusion Proteins, bcr-abl / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D020014:K562 Cells; D016337:RNA, Catalytic / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D016337:RNA, Catalytic; D016044:Fusion Proteins, bcr-abl