基于c-myc启动子的抗肿瘤药物筛选系统的建立

The Construction of a c-myc Promoter-Based Screening System For Anti-Cancer Drugs

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。

To establish a c-myc promoter-based anti-tumor drug screening system, a recombinant plasmid with c-myc promoter driven β- galactosidase reporter gene was constructed and used to create stablely transfected cell lines. Common chemotherapy drugs were used to select the cell lines and to verify their screening specificity and mechanism. Cell based screening assays with this system showed that some common chemotherapy drugs including Doxorubicin, significantly reduced the activity of c-myc promoter and reduce the steady state c-myc mRNA level. In addition, the repressive effect of doxorubicin mediated repression only requires c-myc P2 promoter including a region within the ＋66 nt of c-myc gene. Therefore, this system will be useful not only in the screening for anti-cancer drugs that repress the expression of c-myc oncogene but also in the study of anti-cancer mechanisms of these drugs.