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目的:探讨中性粒细胞活化对肺癌血管生成的影响及其分子机制。方法:以人早幼粒白血病细胞系HL-60为基础,利用DMSO诱导其分化为中性粒细胞样细胞,利用慢病毒感染手段建立低表达PARP-1中性粒细胞系,利用LPS刺激、与肺癌细胞共培养方式活化中性粒细胞,收集中性粒细胞活化上清后进行血管生成实验,应用Realtime-PCR、Western blot和明胶酶谱实验检测血管生成影响因子VEGF和MMP-9的表达情况。结果:中性粒细胞活化产物显著增强肺癌血管生成能力,Western Blot结果显示,低表达PARP-1中性粒细胞构建成功,低表达PARP-1中性粒细胞对肺癌血管生成的促进作用减弱,明胶酶谱实验和Western Blot实验结果显示,MMP-9蛋白的活性及表达量与PARP-1表达量呈负相关,Realtime-PCR结果表明低表达PARP-1中性粒细胞VEGF和MMP-9表达明显减少。结论:中性粒细胞通过PARP-1基因调控MMP-9和VEGF表达促进肺癌血管生成,PARP-1基因可能成为肺癌治疗的新靶点。
Objective: To investigate the effect of neutrophil activation on angiogenesis in lung cancer and its molecular mechanism. Methods: Based on the human promyelocytic leukemia cell line HL-60, the cells were induced to differentiate into neutrophil-like cells by using DMSO. The low expression PARP-1 neutrophil line was established by lentivirus infection. LPS stimulation, Neutrophils were activated by co-culture with lung cancer cells. The neutrophil-activated supernatants were collected for angiogenesis experiments. Realtime-PCR, Western blot and gelatin zymography were used to detect the expression of VEGF and MMP-9 Happening. Results: The activated product of neutrophils significantly enhanced the angiogenesis ability of lung cancer. Western Blot results showed that the low expression of PARP-1 neutrophil was successfully constructed. The low expression of PARP-1 neutrophil decreased the angiogenesis of lung cancer, The results of gelatin zymography and Western Blot showed that there was a negative correlation between the activity of MMP-9 protein and the expression of PARP-1, and Realtime-PCR results showed that the expression of VEGF and MMP-9 in PARP-1 neutrophil obviously decrease. CONCLUSION: PARP-1 gene regulates the expression of MMP-9 and VEGF in neutrophils to promote the angiogenesis of lung cancer. PARP-1 gene may be a new therapeutic target for lung cancer.