顺铂修饰DNA全基因组测序新方法及其在顺铂抗肿瘤分子机制研究中的应用

顺铂是全球三大使用最多的抗肿瘤药物之一，它对包括睾丸癌、卵巢癌等在内的多种癌症有着非常好的治疗效果。顺铂治疗癌症的主要机制是与肿瘤细胞基因组DNA上的碱基发生化学交联，使得DNA复制和转录受到抑制并最终导致细胞死亡。然而，受研究手段的限制，目前对顺铂修饰DNA在全基因组水平上的交联位点却几乎一无所知，严重影响了我们对顺铂作用于基因组并导致细胞死亡的分子机理研究。因此，本项目将开发能够特异性识别并富集含有顺铂修饰的基因组DNA片段新方法，结合高通量测序技术，从而检测顺铂在基因组水平上的分布情况。通过比较对顺铂敏感性不同的癌细胞系中顺铂在基因组上的分布信息，解析顺铂修饰基因组DNA的反应规律，以期在组学水平上加深我们对顺铂抗肿瘤分子机制的理解。

Cisplatin is among the top three most widely used antitumor drugs in the world. It is highly effective in treating testicular and ovarian cancers. The major mechanism of antitumor effect of cisplatin is its ability to crosslink DNA, which inhibits DNA replication and transcription, ultimately leading to cell death. Genomic DNA in the cell nucleus has been shown to be the primary target of cisplatin, yet the crosslinking pattern of cisplatin on the genomic scale remains largely unknown. Such limited knowledge also significantly hinders our study on the molecular mechanism of cisplatin's antitumor effect. Here, we propose a novel chemical biology method in order to profile the interactions between cisplatin and cellular DNA on a genomic scale. Such method involves the development of robust proteins that can specifically recognize and enrich cisplatin-containing DNA, which will then be subjected to high-throughput sequencing. By comparing the cisplatin distribution of cell lines with different cisplatin sensitivity, we aim to uncover the molecular patterns of cisplatin-DNA crosslinking adduct, ultimately leading to a better understanding of the antitumor effect of cisplatin.