肿瘤浸润中性粒细胞在LKB1缺失肺腺癌血管生成中的作用机制研究

LKB1 deficiency is one of the most frequent mutations in lung adenocarcinoma, which is highly enriched in angiogenesis. However, the mechanism is unclear. Previous studies showed that there was more tumor infiltrating neutrophils (TIN) existing in tumor tissues of LKB1-deficient lung cancer mouse model.. Our study found that there was an evaluated level of TIN and tumor microvascular in tumor tissues of lung adenocarcinoma patients with hemoptysis. The level of TIN and LKB1 expression were positively relevant in lung adenocarcinoma tissues. The co-culture of LKB1-deficient adenocarcinoma cells and neutrophils could significantly promote tubule formation, which was inhibited in LKB1-overexpression cells. The co-culture of LKB1-deficient adenocarcinoma cells and neutrophils could also promote epithelial-mesenchymal transition (EMT), which could promote tumor angiogenesis.. Thus, we hypothesize that TIN plays an important role in angiogenesis in LKB1-deficient lung adenocarcinoma. We will establish LKB1-deficient and LKB1-overexpressive cell model and mouse subcutaneous tumor model to investigate the effect of TIN in lung adenocarcinoma angiogenesis. By studying LKB1-deficient lung adenocarcinoma sensitive drugs of TIN to clarify the mechanism of treatment LKB1 missing mechanisms of lung adenocarcinoma, lack of clear TIN in LKB1 has a role in the treatment of lung adenocarcinoma and significance.We aim to explore the mechanism of sensitive drugs of LKB1-deficient lung adenocarcinoma through angiogenesis induced by TIN, and to explore the effect of TIN in treatment of LKB1-deficient lung adenocarcinoma.

LKB1缺失是肺腺癌最常见的突变之一，常伴有血管生成增加，其机制尚不明确。LKB1缺失小鼠肺癌模型中存在更多的肿瘤浸润中性粒细胞(TIN)。我们研究发现：咯血肺腺癌患者的肿瘤组织中存在高水平的TIN及肿瘤微血管；肺腺癌肿瘤组织中TIN与LKB1表达呈明显负相关；LKB1缺失肺腺癌细胞在与中性粒细胞共培养后明显促进了小管形成，而高表达LKB1可以明显抑制该过程；LKB1缺失肺腺癌细胞与中性粒细胞共培养可促进肿瘤细胞上皮间质转化（EMT），后者可促进肿瘤血管生成。. 由此我们推测TIN在LKB1缺失肺腺癌的血管生成中起到重要作用。我们将通过构建LKB1缺失及过表达细胞模型及小鼠皮下种植瘤模型，研究TIN对LKB1缺失肺腺癌细胞血管生成的影响及机制。通过研究LKB1缺失肺腺癌敏感性药物对TIN诱导血管生成的作用，阐明治疗LKB1缺失肺腺癌的新机制，明确TIN在LKB1缺失肺腺癌治疗中的作用及意义。

LKB1缺失是肺腺癌最常见的突变之一，常伴有血管生成增加，其机制尚不明确。LKB1缺失小鼠肺癌模型中存在更多的肿瘤浸润中性粒细胞(TIN)。本课题证明TIN能够促进LKB1缺失肺腺癌血管生成及EMT的发生。同时，本课题找到了与LKB1缺失肺腺癌关系密切的基因FGL1，FGL1与LKB1缺失肺腺癌的血管生成及EMT的发生有密切关系，有望成为治疗LKB1缺失肺腺癌病人的新靶点，同时对LKB1缺失肺腺癌的发展机制进行了补充。也找到了与LKB1缺失肺腺癌密切相关的泛素相关基因SH3RF1，可能成为一个指示LKB1突变肺腺癌病人预后的重要指标，对LKB1突变肺腺癌病人的治疗具有指导意义。..我们利用二代测序、免疫组化等技术在肺腺癌病人肿瘤组织中检测LKB1突变状态及EMT相关指标和血管生成相关指标的表达差异。在人体肺腺癌细胞中，如A549（LKB1缺失型突变的肺腺癌细胞系），过表达LKB1，通过qRT-PCR技术验证两组细胞中LKB1表达状态，并通过qRT-PCR等手段，检测LKB1缺失及野生型细胞中EMT相关指标及血管生成相关指标的表达差异。同时我们也将这两种细胞系，分别与人外周血提取的中性粒细胞共培养，分别利用qRT-PCR等手段检测两组细胞的EMT指标及血管生成指标的表达差异。证明了TIN促进LKB1突变肺腺癌血管生和EMT的发生。..我们利用生物信息学数据挖掘，从TCGA、GEO等生信数据库下载了含有LKB1突变信息肺腺癌病人相关数据，分析找到了与LKB1缺失肺腺癌密切相关的基因FGL1，通过初步实验证明，FGL1低表达能够促进LKB1缺失肺腺癌的血管生成及EMT的发生，FGL1有可能成为治疗LKB1突变肺腺癌的一个新靶点。也找到了与LKB1缺失肺腺癌密切相关的泛素相关基因SH3RF1，SH3RF1可能成为一个指示LKB1突变肺腺癌病人预后的重要指标，对LKB1突变肺腺癌病人的治疗具有指导意义。..本课题从总体上证明TIN能够促进LKB1缺失肺腺癌血管生成及EMT的发生，找到了与LKB1缺失肺腺癌关系密切的基因FGL1与SH3RF1，为 LKB1缺失肺腺癌的发展机制提供了理论补充，也对LKB1突变肺腺癌病人的治疗具有重要指导意义。