Dz13 induces a cytotoxic stress response with upregulation of E2F1 in tumor cells metastasizing to or from bone.
The oligonucleotide Dz13 is a DNA enzyme (deoxyribozyme) that cleaves c-Jun mRNA. It has efficacious effects against tumors directly, is active against tumor-induced angiogenesis, inhibits neointima formation after arterial injury, and controls inflammatory responses. The off-target effects of Dz13 may in fact be driving some of these potentially therapeutic effects, though no mechanisms have been clearly defined in target cells. To this end, we here show that when a panel of human tumor cells that naturally propagate in bone are challenged with Dz13, the tumor suppressor E2F1 is upregulated regardless of cellular p53 status. The piddosomal components, p53-induced protein with a death domain and caspase-2, were translocated to the nucleus when deoxyribozymes were incubated with cells, but RIP associated Ich-1/CED homologous protein with death domain levels increased throughout the cell with either Dz13 or its scrambled control oligonucleotide. In response to Dz13-mediated cytotoxicity, cells upregulated levels of ERK, Akt, and p38. Summarily, these results suggest a cytotoxic stress (resembling DNA damage) response of tumor cells to Dz13, which induces apoptosis via the activation of inhibitor of caspase-activated deoxyribonuclease and protein kinase C delta. In vivo, in tumor-in-bone orthotopic and clinically relevant models for prostate and breast cancer metastasis, and a novel spontaneously metastasizing model for osteosarcoma (OS), Dz13 decreased growth in bone, and also metastasis for OS. This new model for OS was assessed to be clinically relevant in its expression of typical bone markers, osteopontin and osteocalcin. These results provide an off-target mechanism for Dz13 function, but this may be useful therapeutically against tumors.
Dass
Crispin R
CR
Department of Orthopaedics, St. Vincent's Hospital Melbourne, Fitzroy, Australia. cris.dass@yahoo.com
Tan
Mei Lin
ML
Galloway
Stuart J
SJ
Choong
Peter F M
PF
D017209:Apoptosis / Q000187:drug effects*; D001859:Bone Neoplasms / Q000473:pathology* / Q000556:secondary; D045744:Cell Line, Tumor; D004249:DNA Damage; D021881:DNA, Catalytic / Q000494:pharmacology*; D050687:E2F1 Transcription Factor / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D015854:Up-Regulation / Q000187:drug effects*
D021881:DNA, Catalytic; C485475:Dz13 DNAzyme; D050687:E2F1 Transcription Factor; C493753:E2F1 protein, human