ALKYLATING & CLEAVING ANTIC-MYC DNAS FOR BREAST CANCER

烷基化

• 批准号: 6394940
• 负责人: ERIC WICKSTROM
• 金额: $ 3.87万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-07-01 至 2003-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6394940
• 关键词: DNA; MCF7 cell; alkylation; antisense nucleic acid; bleomycin; breast neoplasms; cell line; cell proliferation; chemical cleavage; gene expression; gene therapy; genetic promoter element; laboratory mouse; natural gene amplification; neoplastic cell; nucleic acid sequence; oligonucleotides; protooncogene; tissue /cell culture; tissue mosaicism; xenotransplantation

Breast cancer attacks one out of every nine women. Among the many indicators and prognostic factors in breast cancer, the c- MYC proto-oncogene is amplified and overexpressed in many early and advanced stage mammary tumors. Antisense DNA therapy is proving effective in human clinical trials against viruses and oncogenes. Nuclease resistant reactive oligonucleotides offer powerful tools for intervention against malignant cells with activated oncogenes. The hypothesis of this proposal is that oligonucleotide therapy will achieve antiproliferative potency by specifically ablating c-MYC overexpression in breast cancer cells at subtoxic doses. For this purpose, the Novosibirsk laboratory will synthesize alkylating [4-(N-2-chloroethyl, N-methylamino) benyzylmethylamino] and cleaving (bleomycin A5) derivatives of nuclease resistant phosphorothioate DNAs, 2'-O-methyl RNAs, and their chimeras, potentiated with phenazinium and/or steroid modifications, targeted against c-MYC mRNA targets. The unique heterobifunctional oligonucleotide derivatives with 3'- cholesterol and 5'-reactive groups efficiently penetrate into cells, resist intracellular nucleases, and modify nucleic acids site-specifically. The Novosibirsk laboratory will also synthesize bleomycin A5 derivatives of nuclease resistant triplex-forming DNAs, potentiated with phenazinium and/or steroid modifications, targeted against c-myc DNA oligopurine promoter sequences. The Philadelphia laboratory will test the antigen reduction and antiproliferative efficacy of the reactive oligonucleotides in a tumorigenic human breast cancer cell line, BT474, which overexpresses c-MYC, compared with MCF7 cells, which do not overexpress c-MYC without estrogen stimulation. This new joint effort has been initiated by the visit of Dr. Smith to the Novosibirsk laboratory to measure anti-c-MYC bleomycin A5 oligomer reactivity with c-MYC RNA. In future years, the most potent nuclease resistant reactive oligonucleotides determined in cell culture will then be tested for preclinical potency by measuring inhibition of tumor growth in nude mice due to subcutaneous implantation of the BT474 and MCF7 cell lines.

乳腺癌攻击每9名妇女中的一个。 在乳腺癌的许多指标和预后因素中，C- MYC原始癌基因在许多早期和晚期乳腺肿瘤中被放大和过表达。 反义DNA疗法在针对病毒和癌基因的人类临床试验中有效。 抗核酸酶的反应性寡核苷酸为用激活的癌基因对恶性细胞进行干预提供了强大的工具。 该提议的假设是寡核苷酸疗法将通过特异性地烧蚀乳腺癌细胞中的C-Myc过表达来实现抗增殖效力，这是在乳腺癌细胞中的。 For this purpose, the Novosibirsk laboratory will synthesize alkylating [4-(N-2-chloroethyl, N-methylamino) benyzylmethylamino] and cleaving (bleomycin A5) derivatives of nuclease resistant phosphorothioate DNAs, 2'-O-methyl RNAs, and their chimeras, potentiated with phenazinium and/or steroid针对C-MYC mRNA靶标的修饰。 具有3'-胆固醇和5'反应组的独特异质核苷酸衍生物有效地渗透到细胞中，抗​​细胞内核酸酶，并特异性地修饰核酸。 Novosibirsk实验室还将合成抗核酸酶抗三酶形成的DNA的博来霉素A5衍生物，该DNA具有苯并依和/或类固醇修饰的增强，针对C-MYC DNA少肽氨酸启动子序列。 费城实验室将测试反应性寡核苷酸在肿瘤性人类乳腺癌细胞系BT474中的抗原降低和抗增殖功效，与MCF7相比，它过表达C-MYC与MCF7细胞相比，该细胞过表达C-MYC，而MCF7细胞不会过度表现C-Myc，而无需过表达C-Myc。史密斯博士对Novosibirsk实验室的访问开始了这一新的联合努力，以测量与C-Myc RNA的抗C-Myc Bleyomycin A5低聚物反应性。 在未来的几年中，通过测量因BT474和MCF7细胞系的皮下植入而测量裸鼠肿瘤生长的抑制，将测试在细胞培养中确定的最有效的核酸酶反应性寡核苷酸。