基于前期GWAS芯片的宫颈癌化疗疗效相关SNPs位点筛选与功能研究

In the past 20 years, although neoadjuvant chemotherapy has not yet became the standard treatment modality in cervical cancer, it was widely used before radical surgery to shrink the tumor lesion size, reduce micro-metastases, increase surgical resection rate and improve the prognosis in cervical cancer in many countries. At present, the effect of NACT on long-term prognosis remains controversial, but it is clear that the effect of NACT on long-term prognosis is closely related to its clinical response. Therefore choosing NACT or not individually by prediction of chemotherapy response is of great significance to improve the overall survival rate. Because there are significant individual differences of clinical response to NACT, and we investigate its relationship with the germline polymorphism in the genomic DNA. Previously, we conducted a genome-wide association study (GWAS) of cervical cancer in the Han Chinese population. Based on the data, we have conducted a genome-wide examination of novel genetic variants associated with response to platinum-based chemotherapy in cervical cancer patients treated with neoadjuvant chemotherapy and radical surgery. And we have identified several strong association between clinical response and 25 SNPs (p ≤ 1×10-4). We use time-of-flight mass spectrometry genotyping technology in a large sample set, and validate the SNPs closely related to neoadjuvant chemotherapy. Through bioinformatics analysis and molecular functional analysis, on one hand, an effective and viable solution to individualized therapy of neoadjuvant chemotherapy for cervical cancer patients can be provided in our study. On the other hand, we can investigate the mechanism of chemotherapy in cervical cancer, and improve the treatment and the prognosis , which provide a theoretical basis for further research.

近20多年来，新辅助化疗（NACT）虽不是宫颈癌标准治疗模式，但因其缩小肿瘤大小、减少微转移灶、提高手术切除率，在我国等很多国家被广泛应用。目前，NACT对于远期预后的作用仍存在争议，而比较明确的是NACT远期预后作用与化疗短期疗效息息相关。因此通过预测化疗反应性，对患者进行个体化选择对于改善总体生存率有重要的意义。因化疗疗效存在明显个体差异，本课题从胚系基因组DNA遗传多态性这一最能体现个体差异性的角度出发，将前期宫颈癌全基因组SNP分型芯片重新入组分析，已成功筛选到与NACT疗效相关的25个SNPs位点（p ≤ 1×10-4），本项目拟在大样本中采用飞行时间质谱基因分型技术进行分型验证，并对验证得到的SNP位点进行生物信息学分析及分子生物学功能研究，一方面，为广大地区NACT个体化选择提供参考；另一方面，为进一步了解宫颈癌化疗作用机制、提高化疗治疗水平、改善预后提供重要的理论基础。

目前，新辅助化疗（NACT）虽不是宫颈癌标准治疗模式，但因其缩小肿瘤大小、减少微转移灶、提高手术切除率，在我国、韩国、日本等很多国家被广泛应用。目前，NACT对于远期预后的作用仍存在争议，而比较明确的是NACT远期预后作用与化疗短期疗效息息相关。因此通过预测化疗反应性，对患者进行个体化选择对于改善总体生存率有重要的意义。本课题从胚系基因组DNA遗传多态性这一最能体现个体差异性的角度出发，将前期宫颈癌全基因组SNP分型芯片重新入组分析，对 226例样本进行了全基因组关联分析。通过 160例新辅助化疗敏感性患者和 66例新辅助化疗不敏感患者的比较，得到 27个 SNP 的 P<1×10 -5，并挑选了 23个 SNP 位点进行两轮验证，然后将芯片部分和两轮验证结果进行meta分析，结果发现有4个SNP位点的P值<5.0×10 -5。其中rs6812281的 P 值为 9.0×10 -9 ，rs1742101和 rs1364121的 P 值分别为 7.11×10 -6 和 3.15×10 -6，rs4590782的 P值分别为 1.59×10 -5。且我们发现rs4590782与预后密切相关：总生存率比较(82.9% vs. 75.8%, P = 0.083)、无瘤生存率比较（80.8% vs. 72.7%, P = 0.021）。本项目所得到的研究结果为NACT个体化选择提供参考；并为进一步了解宫颈癌化疗作用机制、提高化疗治疗水平、改善预后提供重要的理论基础。