Antisense TRPM-2 oligodeoxynucleotides chemosensitize human androgen-independent PC-3 prostate cancer cells both in vitro and in vivo.
Although numerous chemotherapeutic regimens have been evaluated for patients with hormone-refractory prostate cancer, none has improved survival. Testosterone-repressed prostate message-2 (TRPM-2), which is highly up-regulated after androgen withdrawal and during androgen-independent progression in prostate cancer, has been shown to inhibit apoptosis induced by various kinds of stimuli. The objectives in this study were to test whether antisense (AS) oligodeoxynucleotides (ODNs) targeted against TRPM-2 enhance chemosensitivity in human androgen-independent prostate cancer PC-3 cells both in vitro and in vivo. Initially, the potency of 10 AS ODNs targeting various regions of the TRPM-2 mRNA were evaluated, and the AS ODN targeted to the TRPM-2 translation initiation site (AS ODN#2) was found to be the most potent sequence for inhibiting TRPM-2 expression in PC-3 cells. Despite significant dose-dependent and sequence-specific suppression of TRPM-2 expression, AS ODN#2 had no effect on growth of PC-3 cells both in vitro and in vivo. However, pretreatment of PC-3 cells with AS ODN#2 significantly enhanced chemosensitivity of Taxol (paclitaxel) and mitoxantrone in vitro. Characteristic apoptotic DNA laddering and cleavage of poly(ADP-ribose) polymerase were observed after combined treatment with AS ODN#2 plus paclitaxel or mitoxantrone but not with either agent alone. In vivo administration of AS ODN#2 plus either paclitaxel or mitoxantrone significantly decreased PC-3 tumor volume by 80 or 60%, respectively, compared with mismatch control ODN plus either paclitaxel or mitoxantrone. In addition, terminal deoxynucleotidyl transferase-mediated nick end labeling staining revealed increased apoptotic cells in tumors treated with AS ODN#2 plus paclitaxel or mitoxantrone. These findings confirm that TRPM-2 overexpression confers resistance to cytotoxic chemotherapy in prostate cancer cells and illustrates the potential utility of combined treatment with AS TRPM-2 ODN plus chemotherapeutic agents for patients with hormone-refractory prostate cancer.
Miyake
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The Prostate Centre, Vancouver General Hospital, British Columbia, Canada.
Chi
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Gleave
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ME
D000728:Androgens / Q000502:physiology; D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D002455:Cell Division / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D051152:Clusterin; D053938:DNA Fragmentation / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D004357:Drug Synergism; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006023:Glycoproteins / Q000235:genetics*; D006801:Humans; D020287:In Situ Nick-End Labeling; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D008942:Mitoxantrone / Q000494:pharmacology; D018832:Molecular Chaperones; D020319:Oligodeoxyribonucleotides, Antisense / Q000302:isolation & purification / Q000494:pharmacology* / Q000627:therapeutic use; D017239:Paclitaxel / Q000494:pharmacology; D011471:Prostatic Neoplasms / Q000473:pathology / Q000628:therapy*; D012333:RNA, Messenger / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D013873:Thionucleotides / Q000302:isolation & purification / Q000494:pharmacology / Q000627:therapeutic use; D014407:Tumor Cells, Cultured
D000728:Androgens; D000970:Antineoplastic Agents; C495242:CLU protein, human; C495243:Clu protein, mouse; D051152:Clusterin; D006023:Glycoproteins; D018832:Molecular Chaperones; D020319:Oligodeoxyribonucleotides, Antisense; D012333:RNA, Messenger; D013873:Thionucleotides; D008942:Mitoxantrone; D017239:Paclitaxel