MiR-9调控区域遗传变异与非小细胞肺癌预后的关联及其生物学功能的研究

The five-year survival rate of lung cancer is less than 15% and characterized by individual variations. There have been so far very few reported about molecular markers that are able to evaluate prognosis of lung cancer. Studies have indicated that miRNA is involved in tumor development through regulating target genes. However, its single-nucleotide polymorphisms associated with lung cancer prognosis has been very few reported yet. We investigated the correlation between 88 important miRNA SNPs and prognosis of lung caner. The genotyping was performed by Illumina Infinium® BeadChip (Illumina Inc.). As a result, we found that the rs1501672 A>G, which is in the upstream of miR-9, demonstrated relatively strong correlation with the prognosis of lung caner. Based on the previous study, we are going to carry out the work as the following: First, Verify the changes of the expression of the miRNA are resulted from the above SNP’s modifications to the binding between the transcription factor and the promoter based on the EMSA. Second, explore further the regulating mechanism of the development of lung cancer prognosis affected by miR-9 SNPs regulating target gene. Eventually, the study will illuminate the relations and possible mechanisms between miR-9 SNPs and lung cancer prognosis, and provide theoretical evidences to the relevant markers of becoming the individualized evaluation for lung cancer prognosis and innovative ideas to the clinical individual treatments.

肺癌的5年生存率低于15%,且存在明显的个体差异,迄今能个体化评估肺癌预后的分子标志物甚少。研究表明miRNA通过对靶基因的调控参与了肿瘤的发生发展，但其调控区域SNPs与非小细胞肺癌预后的关联研究鲜有报道。我们利用外显子芯片技术通过1001例大样本病例研究，分析88个与肿瘤相关的重要miRNA调控区域SNPs与非小细胞肺癌预后的关联，结果发现位于miR-9上游的rs1501672 A>G与肺癌较好的预后相关。本课题拟在前期研究基础上通过①EMSA等方法来验证rs1501672 A>G通过改变转录因子HSF2与启动序列结合的能力，从而导致miR-9表达量的变化；②通过体内外实验探索miR-9通过对靶基因CDH1的调节影响肺癌发展预后的调控机制。最终阐明miR-9 SNPs与肺癌预后的关联及可能的生物学机制，为其成为个体化评估肺癌预后的分子标记物提供理论依据,为肺癌个体化治疗提供新的思路.

肺癌的5年生存率低于15%，且存在明显的个体差异，迄今能个体化评估肺癌预后的分子标志物甚少。研究表明miRNA通过对靶基因的调控参与了肿瘤的发生发展，但其调控区域SNPs与非小细胞肺癌预后的关联研究鲜有报道。我们利用外显子芯片技术通过1001例大样本病例研究，分析88个与肿瘤相关的重要miRNA调控区域SNPs与非小细胞肺癌预后的关联，结果发现位于miR-9上游的rs1501672(A>G)与肺癌较好的预后相关，位于miR-125b-1上游的rs2241490(G>A)与肺癌较差的预后相关，利用双荧光素酶报告基因发现位于miR-9-2上游调控区域的rs1501672(A>G)基因型改变可能影响了转录因子与miR-9-2上游启动序列的结合，从而改变了成熟miR-9量的表达，位于miR-125b-1上游的rs2241490(G>A)基因型改变潜在影响了转录因子与miR-125b上游序列结合改变了miR-125b的表达量。我们进一步利用生物信息学分析发现miR-9在肺癌组织中呈高表达，并绘制了以miR-9为中心的CeRNA网络，通过进一步分析发现潜在和miR-9结合的靶基因LIFR，及两条LncRNA（GVINP1、KIAA0087）在肺癌组织中表达量下降，与miR-9表达量负相关，而且LIFR、GVINP1、KIAA0087表达量升高与肺癌病人较好的预后相关；最后通过体内外实验进一步探讨miR-9表达量的失调可以影响肺癌细胞的增殖和侵袭能力，并在肺癌的发展及预后过程中的发挥作用。通过上述实验最终阐明microRNA SNPs与肺癌预后的关联及可能的生物学机制，为其成为个体化评估肺癌预后的分子标记物提供理论依据,为肺癌个体化治疗提供新的思路.