Establishment of a new gene therapy for cancer using antisense ODG and atelocollagen

背景和目的：大量证据支持鸟嘌呤脱羧酶（ODC）在人类肿瘤发生和维持中的直接作用。虽然针对不同基因的反义寡核苷酸疗法对癌症治疗是有用的，但主要的限制之一是递送问题。我们在这里描述了一种新的反义寡核苷酸递送方法，该方法允许使用间胶原蛋白在体内延长ODC反义寡核苷酸的维持和释放。方法：采用MTT法研究胶原间质中ODC反义寡核苷酸对胃肠道肿瘤（mkn45、COLO201）和横纹肌肉瘤（RD）细胞生长的影响。在体内，研究了单次给药ODC反义寡核苷酸对MKN45、COLO201和RD细胞肿瘤生长的影响。测定ODC活性和多胺含量。结果：ODC反义寡核苷酸在体外可显著抑制MKN45、COLO201和RD细胞的生长。在3542天的时间内，通过三种途径在胶原间质中单次给予反义寡核苷酸，显著抑制了MKN45、COLO201和RD肿瘤的生长。结论：ODC在多种人类肿瘤中均有显著表达，提示这种新的反义方法可能对人类癌症的治疗具有相当的价值。

Background and Aims: Substantial evidence supports a direct role of ornitine decarboxylase (ODC) in the development and maintenance of human tumors. Although antisenseoligonucleotide therapy targeting various genes are useful for cancer treatment, one of the major limitations is the problem of delivery. We describe here a novel antisense oligonucleotide delivery method which allows prolonged sustainment and release of ODC antisense oligonucleotides in vivo using atelocollagen. Methods: The effect of ODC antisense oligonucleotides in the atelocollagen on cell growth of gastrointestinal cancer (MKN 45 and COLO201), and rhabdomyosarcoma (RD) was studied in vitro using MTT assay. In vivo, the effect of intratumoral, intramuscular and intraperitoneal single administration of ODC antisense oligonucleotides in the atelocollagen on tumor growth of MKN45, COLO201 and RD cells was examined. ODC activity and polyamine contents were measured. Results: In vitro, ODC antisense oligonucleotides in the atelocollagen remarkably suppressed MKN45, COLO201 and RD cell growth. A single administration of antisense oligonucleotides in the atelocollagen via three routes remarkably suppressed the growth of MKN45, COLO201 and RD tumor over a period of 3542 days. Conclusion: As various human cancers significantly express ODC, the results strongly suggest that this new antisense method may be of considerable value for treatment of human cancers.