论文部分内容阅读
建立基于c-myc启动子,以β-半乳糖苷酶报告基因(β-galactosidase reporter gene)的新型抗肿瘤药物筛选系统。构建含有c-myc启动子和β-半乳糖苷酶报告基因的重组质粒,转染U2OS细胞,获取稳定表达β-半乳糖苷酶的细胞株并使用常用抗肿瘤药物对其筛选特异性与筛选机制进行鉴定。细胞水平常用化疗药物的筛选验证实验表明,部分临床化疗药物如阿霉素在这一系统中能显著下调c-myc启动子的活性和c-myc mRNA的量,同时证实阿霉素对c-myc启动子的作用位点在+66位以前的P2启动子。因而通过建立基于c-myc启动子的抗肿瘤药物筛选系统,不但可以有效筛选能够抑制c-myc表达的抗肿瘤药物,也可以研究这类药物的作用机理。
A novel antitumor drug screening system based on c-myc promoter and β-galactosidase reporter gene was established. The recombinant plasmid containing c-myc promoter and β-galactosidase reporter gene was constructed and transfected into U2OS cells to obtain the cell strain stably expressing β-galactosidase and its screening specificity and screening using common anti-tumor drugs Mechanism for identification. Cell-level screening of commonly used chemotherapeutic drugs validation experiments showed that some of the clinical chemotherapy drugs such as doxorubicin in this system can significantly reduce the c-myc promoter activity and the amount of c-myc mRNA at the same time confirmed doxorubicin c- The P2 promoter, where the role of the myc promoter is located before +66. Therefore, by establishing a c-myc promoter-based antitumor drug screening system, not only can effectively screen anti-tumor drugs that can inhibit c-myc expression, but also can study the mechanism of action of such drugs.