Adenovirus-mediated expression of both antisense ornithine decarboxylase and s-adenosylmethionine decarboxylase induces G1 arrest in HT-29 cells.
To evaluated the effect of recombinant adenovirus Ad-ODC-AdoMetDCas which can simultaneously express both antisense ornithine decarboxylase (ODC) and Sadenosylmethionine decarboxylase (AdoMetDC) on cell cycle distribution in colorectal cancer cell and investigated underlying regulatory responses, human colorectal cancer cells HT-29 were cultured in RPMI 1640 medium and infected with Ad-ODC-AdoMetDCas. Cell cycle progression was detected by flow cytometry analysis. The expression levels of cell cycle regulated proteins were measured by Western blot analysis. The mRNA level of cyclin D1 was measured by RT-PCR. And a luciferase reporter plasmid of cyclin D1 promoter was constructed to observe the effect of Ad-ODC-AdoMetDCas on cyclin D1 promoter activity. The results showed that recombinant adenovirus Ad-ODC-AdoMetDCas significantly induced G1 arrest, decreased levels of cyclin D1 protein and mRNA and suppressed the promoter activity. Ad-ODC-AdoMetDCas also inhibited nuclear translocation of beta-catenin. In conclusion, downregulation of ODC and AdoMetDC mediated by Ad-ODC-AdoMetDCas transfection induces G(1) arrest in HT-29 cells and the arrest was associated with suppression of cyclin D1 expression and inhibition of beta-catenin nuclear translocation. As a new anticancer reagent, the recombinant adenovirus Ad-ODC-AdoMetDCas holds promising hope for the therapy of colorectal cancers.
Gong
Lei
L
Institute of Biochemistry and Molecular Biology, School of Medicine, Shandong University, Jinan 250012, China.
Jiang
Chunying
C
Zhang
Bing
B
Hu
Haiyan
H
Wang
Wei
W
Liu
Xianxi
X
D012437:Adenosylmethionine Decarboxylase / Q000235:genetics*; D000256:Adenoviridae / Q000235:genetics; D002453:Cell Cycle / Q000187:drug effects; D019938:Cyclin D1 / Q000235:genetics / Q000378:metabolism; D016193:G1 Phase / Q000187:drug effects*; D015870:Gene Expression / Q000502:physiology*; D017930:Genes, Reporter; D005822:Genetic Vectors; D019073:HT29 Cells; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D020319:Oligodeoxyribonucleotides, Antisense / Q000096:biosynthesis / Q000494:pharmacology*; D009955:Ornithine Decarboxylase / Q000235:genetics*; D051176:beta Catenin / Q000378:metabolism
D020319:Oligodeoxyribonucleotides, Antisense; D051176:beta Catenin; D019938:Cyclin D1; D008156:Luciferases; D009955:Ornithine Decarboxylase; D012437:Adenosylmethionine Decarboxylase