Effects of antisense RNA targeting of ODC and AdoMetDC on the synthesis of polyamine synthesis and cell growth in prostate cancer cells using a prostatic androgen-dependent promoter in adenovirus.
OBJECTIVE
This study was designed to investigate the use of a prostatic androgen-dependent promoter to mediate antisense targeting of ornithine decarboxylase (ODC) and S-adenosylmethionine decarboxylase (AdoMetDC) and its effects on the synthesis of polyamine. We also examined the potential of this construct for prostate cancer therapy.


METHODS
pADxsi-PSES-AdoMetDC-ODC-PolyA AV was constructed and used to infect various cancer cell lines, including LNCaP, HT-29, H1299, HepG2. The effects of pADxsi-PSES-AdoMetDC-ODC-PolyA AV on the expression of ODC and AdoMetDC, in addition to the cell cycle, apoptosis and p21 levels, were analyzed through Western blotting and cytometry. A Matrigel invasion assay was used to analyze the effects of the recombinant virus on tumor cell invasion. The effect on polyamine content was also determined, and the relationship between inhibition of cellular ODC and AdoMetDC and decreases in polyamine were also investigated using a polyamine recovery assay.


RESULTS
Treatment with pADxsi-PSES-AdoMetDC-ODC-PolyA at an MOI of 90 significantly inhibited the proliferation of LNCaP cells, which could not be recovered through the addition of exogenous putrescine. The expression of ODC and AdoMetDC was also reduced, as was the polyamine content. The G1 phase of LNCaP cells was delayed, but no increase in apoptosis was detected. The down-regulation of ODC and AdoMetDC led to increased p21 expression.


CONCLUSIONS
The pADxsi-PSES-AdoMetDC-ODC-PolyA AV specifically inhibited the expression of ODC and AdoMetDC and the synthesis of polyamine, while it induced p21 expression, resulting in cell growth arrest in the G1 phase in prostate cancer cells but not in other cells.
Li
Wei
W
Institute of Biochemistry and Molecular Biology, School of Medicine, Shandong University, Jinan, Shandong, China.
Liu
Xianxi
X
Wang
Wei
W
Sun
Hui
H
Hu
Yongmei
Y
Lei
Hong
H
Liu
Guanghui
G
Gao
Yanyan
Y
D000230:Adenocarcinoma / Q000378:metabolism* / Q000473:pathology; D012437:Adenosylmethionine Decarboxylase / Q000235:genetics / Q000378:metabolism*; D000256:Adenoviridae / Q000235:genetics; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects*; D050759:Cyclin-Dependent Kinase Inhibitor p21 / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D008297:Male; D009955:Ornithine Decarboxylase / Q000235:genetics / Q000378:metabolism*; D011073:Polyamines / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics; D011471:Prostatic Neoplasms / Q000378:metabolism* / Q000473:pathology; D016372:RNA, Antisense / Q000235:genetics / Q000494:pharmacology*
D050759:Cyclin-Dependent Kinase Inhibitor p21; D011073:Polyamines; D016372:RNA, Antisense; D049452:Green Fluorescent Proteins; D009955:Ornithine Decarboxylase; D012437:Adenosylmethionine Decarboxylase