Experimental cancer gene therapy by multiple anti-survivin hammerhead ribozymes.
To improve the efficacy of gene therapy for cancer, we designed four hammerhead ribozyme adenoviruses (R1 to R4) targeting the exposed regions of survivin mRNA. In addition to the in vitro characterization, which included a determination of the sequence specificity of cleavage by primer extension, assays for cell proliferation and for in vivo tumor growth were used to score for ribozyme efficiency. The resulting suppression of survivin expression induced mitotic catastrophe and cell death via the caspase-3-dependent pathway. Importantly, administration of the ribozyme adenoviruses inhibited tumor growth in a hepatocellular carcinoma xenograft mouse model. Co-expression of R1, R3 and R4 ribozymes synergistically suppressed survivin and, as this combination targets all major forms of the survivin transcripts, produced the most potent anti-cancer effects. The adenoviruses carrying the multiple hammerhead ribozymes described in this report offered a robust gene therapy strategy against cancer.
Fei
Qi
Q
Fudan University School of Medicine, Shanghai 200032, China.
Zhang
Hongyu
H
Fu
Lili
L
Dai
Xinlan
X
Gao
Baomei
B
Ni
Min
M
Ge
Chao
C
Li
Jinjun
J
Ding
Xia
X
Ke
Yuwen
Y
Yao
Xuebiao
X
Zhu
Jingde
J
D000256:Adenoviridae / Q000235:genetics; D017209:Apoptosis; D006528:Carcinoma, Hepatocellular / Q000235:genetics* / Q000473:pathology* / Q000628:therapy; D045744:Cell Line, Tumor; D002470:Cell Survival; D020868:Gene Silencing; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins; D008869:Microtubule-Associated Proteins / Q000235:genetics*; D009363:Neoplasm Proteins / Q000235:genetics*; D016337:RNA, Catalytic / Q000235:genetics* / Q000627:therapeutic use*; D000077022:Survivin; D014162:Transfection / Q000379:methods*; D016896:Treatment Outcome
C107489:BIRC5 protein, human; D051034:Inhibitor of Apoptosis Proteins; D008869:Microtubule-Associated Proteins; D009363:Neoplasm Proteins; D016337:RNA, Catalytic; D000077022:Survivin; C427204:hammerhead ribozyme