DNAzyme-mediated cleavage of survivin mRNA and inhibition of the growth of PANC-1 cells.
OBJECTIVE
Survivin has functions in both the regulation of cell division and the inhibition of apoptosis, and it is expressed in most human tumors but not in normal adult tissues. It is a potential target for cancer gene therapy. In the present study, we designed a DNAzyme targeting human survivin mRNA and tried to determine its effect on human pancreatic carcinoma cell line PANC-1.


METHODS
We designed and synthesized a '10-23' antisurvivin mRNA DNAzyme, testified its cleavage activity by cell-free test, then delivered it into the PANC-1 cell line through liposome, and detected its effect on survivin mRNA expression by reverse transcription polymerase chain reaction, Western blot methods, evaluated its influence on the growth of PANC-1 cells by apoptosis detection, growth curve, and flow cytometry.


RESULTS
Our results showed that the DNAzyme digested mRNA substrates of survivin efficiently in a dosage- and time-dependent manner, markedly increased apoptosis and inhibited the growth of human pancreatic carcinoma cell line PANC-1, compared with the disabled DNAzyme and untreated controls.


CONCLUSIONS
Our results showed that the designed DNAzyme against survivin mRNA is a good candidate for cancer gene therapy of human pancreatic carcinoma.
Liang
Zhiyong
Z
Department of Pathology, Peking Union Medical College Hospital, Chinese Academy of Medical Science, Beijing, China.
Wei
Shuanzeng
S
Guan
Jian
J
Luo
Yufeng
Y
Gao
Jie
J
Zhu
Hong
H
Wu
Shafei
S
Liu
Tonghua
T
D017209:Apoptosis / Q000187:drug effects; D015153:Blotting, Western; D002277:Carcinoma / Q000473:pathology*; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D021881:DNA, Catalytic / Q000008:administration & dosage / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins; D008869:Microtubule-Associated Proteins / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D009363:Neoplasm Proteins / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D010190:Pancreatic Neoplasms / Q000378:metabolism / Q000473:pathology* / Q000503:physiopathology*; D012333:RNA, Messenger / Q000187:drug effects*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000077022:Survivin; D013997:Time Factors
C107489:BIRC5 protein, human; D021881:DNA, Catalytic; D051034:Inhibitor of Apoptosis Proteins; D008869:Microtubule-Associated Proteins; D009363:Neoplasm Proteins; D012333:RNA, Messenger; D000077022:Survivin