基因多态位点预测云南非小细胞肺癌预后风险研究

Lung cancer is one of the leading causes of the cancer-related mortality in China. The 5-year overall survival rate remains at only 16% for none small cell lung cancer (NSCLC). After surgery, the early-stage cancer patients could receive treatment only based on the TNM stages, which may cause excessive or inadequate treatment in clinical practice. So far, the prediction methods for prognostic risk of these patients have not been established at genetic level. In this situation, we take the advantage of the sample resources in the high NSCLC incidence area at Yunnan province to carry out a retrospective cohort study. The paraffin embedded specimens from 300 patients with NSCLC were collected in five years, and these patients were followed up. In order to reveal difference of their susceptibility loci and genetic diversity, the 44 single nucleotide polymorphisms (SNPs) from genome-wide association study identified in the literatures and their haplotypes will be tested and evaluated, and the frequency and distribution of SNPs in the same sites among different groups will be discussed. These SNPs will be assessed by the survival analysis system and the Cox proportional hazard modeling. The prediction methods of prognostic risk for the patients with stage I or stage II - NSCLC in Yunnan province will be established and it will be a new method of detecting a combination of multiple genes. These findings may be helpful in guiding the clinical practice to design postoperative treatment protocols for the early stage lung cancer patients, to improve diagnostic efficiency and therapeutic efficacy, and to prolong the patients' survival time.

中国肺癌死亡率已居各种肿瘤的首位。非小细胞肺癌患者五年生存率仅为16%。早期术后患者仅依据TNM分期进行治疗存在治疗不足或过度治疗的风险，而迄今从基因水平对这类患者进行预后风险预测方法尚未建立。鉴于此，本项目将充分利用云南肺癌高发的样本资源优势，采用回顾性队列研究方法，收集近5年云南非小细胞肺癌病理石蜡切片300份，并对患者进行随访；针对文献报道通过全基因组关联分析获得的44个多态位点进行基因分型及单倍型构建，比较不同人群间相同基因位点数据，揭示其易感位点的差异及遗传特征；利用生存分析系统及Cox风险评估系统对上述SNPs位点进行评估, 建立针对云南非小细胞肺癌I期、II期患者预后风险预测的多基因联合检测新方法，有助于制定早期肺癌患者术后治疗最佳方案，提高诊断效率、治疗效应及患者生存期, 具有临床指导意义。

非小细胞肺癌(NSCLC)是一种常见恶性肿瘤，迄今从基因水平对早期术后患者进行预后风险预测的方法尚未建立。项目组利用云南肺癌高发的样本资源优势，采集NSCLC石蜡包埋切片、新鲜组织和血液样本，开展流行病学资料收集、整理及分析，并对患者进行随访。对全基因组关联分析获得的44个多态位点进行基因分型及单倍型构建，发现仅有基因EYA2和STK39多态位点与NSCLC发生相关。EYA2基因7个位点基因型（rs11699842AA、rs13039645TT、rs1150441TT、rs1150442CT/CC、rs13041757GA/AA、rs11697925GG、rs6066146CC）增加患病风险，而3个位点基因型（rs6066146CT、rs11696413AA/AC、rs13041611CT）降低患病风险；单倍型ATCCAAATCGCTG、TCTTCAACTGCTG和TCTTCAATTGCTA增加患病风险，而单倍型TCCCCGATCGCTG、TCCTCGACCGCTG和TCCTCGATCGCTG降低风险。STK39基因rs1829227AA/GA基因型增加患病风险,而rs10198473CT/TT降低患病风险； ATTT单倍型增加患病风险。.根据国际疾病基因研究最新趋势，在完成44个多态位点基因分型及单倍型构建的同时，增加了基因功能研究发现：1、EYA2蛋白高表达具有促癌作用，其CpG岛甲基化状态与miR-30a表达情况均影响EYA2蛋白表达水平，继而影响NSCLC细胞增殖、迁移和侵袭等生物学行为。2、GCNT3表达显著上调，敲低GCNT3后显著抑制了NSCLC细胞的增殖、迁移和侵袭能力，阻滞细胞周期进程，促进细胞凋亡。3、miRNA-320a在肝癌组织表达水平下调，而长链非编码RNA-AK091631在大肠癌组织的表达水平明显下调；miR-320a能通过调控c-Myc表达从而有效降低肝癌细胞系的增殖和侵袭能力，而RNA-AK091631作为抑癌基因可以通过调控WNT/β-catenin信号通路从而影响结直肠癌的发生。.本项目通过关联分析与机制探索相结合的研究策略，得到与NSCLC发生相关的3个易感基因，有可能成为NSCLC的诊疗和预后检测的新指标。项目成果发表SCI论文4篇，在投2篇；3人获博士学位、7人获硕士学位。