Targeted expression of Escherichia coli purine nucleoside phosphorylase and Fludara® for prostate cancer therapy.
BACKGROUND
Previous studies have shown that Herpes Simplex Virus thymidine kinase (HSV-tk)/ganciclovir (GCV) comprised the most commonly used suicide gene therapy for prostate cancer, with modest results being obtained. However, novel suicide genes, such as Escherichia coli purine nucleoside phosphorylase (PNP), have been utilized to demonstrate more potent tumor killing and an enhanced bystander effect on local, non-expressing cells compared to HSV-tk.


METHODS
PNP/fludarabine (Fludara®; fludarabine phosphate; Berlex Labs, Richmond, CA, USA) was deliveried by prostate-specific, rat probasin-based promoter, ARR2PB. After infection of various cell lines with ADV.ARR(2) PB-PNP and administration of androgen analog, R1881, expression of PNP mRNA was detected; in vivo, the antitumor effect of the ARR(2) PB-PNP/Fludara system was monitored and analyzed, as well as animal survival.


RESULTS
After in vitro infection with ADV.ARR(2) PB-PNP (multiplicity of infection  =  10), LNCaP cells were more sensitive to a lower concentration Fludara (LD(50) , approximately 0.1 µg/ml) in the presence of R1881. Furthermore, robust bystander effects after R1881/Fludara treatment were observed in LNCaP cells after infection with bicistronic vector ADV.ARR2PB/PNP-IRES-EGFP in contrast to a much weaker effect in cells treated with ADV.CMV-HSV-tk/GCV. In vivo, tumor size in the ADV.ARR2PB-PNP/Fludara treatment group was dramatically smaller than in the control groups, and the mice treated with our system had a significantly prolonged survival, with three of eight mice surviving up to the 160-day termination point, as well as no systemic toxicity.


CONCLUSIONS
The ARR(2) PB-PNP/Fludara system induced massive tumor cell death and a prolonged life span without systemic cytotoxicity; therefore, it might be a more attractive strategy for suicide gene therapy of prostate cancer.
State Key Laboratory of Oncology in South China, Sun Yat-Sen University Cancer Center, Guangzhou, China; Department of Breast Oncology, Sun Yat-Sen University Cancer Center, Guangzhou, China.
Xinhua
X
Xie
Jiaoli
J
Guo
Yanan
Y
Kong
Gene Xianquan
GX
Xie
Laishen
L
Li
Ning
N
Lv
Xiangsheng
X
Xiao
Jun
J
Tang
Xi
X
Wang
Peng
P
Liu
Mingtian
M
Yang
Zeming
Z
Xie
Weidong
W
Wei
David M
DM
Spencer
Xiaoming
X
Xie
D000818:Animals; D019390:Arrestins / Q000235:genetics; D016923:Cell Death / Q000187:drug effects / Q000235:genetics; D045744:Cell Line, Tumor; D004926:Escherichia coli; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects / Q000235:genetics; D041722:Genes, Transgenic, Suicide / Q000235:genetics; D015316:Genetic Therapy; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D008297:Male; D015741:Metribolone / Q000008:administration & dosage; D051379:Mice; D011401:Promoter Regions, Genetic; D011471:Prostatic Neoplasms / Q000235:genetics / Q000473:pathology / Q000628:therapy; D011683:Purine-Nucleoside Phosphorylase / Q000235:genetics* / Q000627:therapeutic use; D051381:Rats; D001084:Vidarabine Phosphate / Q000031:analogs & derivatives* / Q000627:therapeutic use; D000071557:beta-Arrestins
D019390:Arrestins; D000071557:beta-Arrestins; C485184:enhanced green fluorescent protein; D001084:Vidarabine Phosphate; D049452:Green Fluorescent Proteins; C042382:fludarabine phosphate; D015741:Metribolone; D011683:Purine-Nucleoside Phosphorylase