Chen Kai, Li Yajie, Xu Hui, Zhang Chunfeng, Li Zhiqiang, Wang Wei, Wang Baofeng
Department of Respiratory Medicine, Baoji Central Hospital, Baoji 721008, Shaanxi, China.
Department of Cardiology, Baoji Central Hospital, Baoji 721008, Shaanxi, China.
Oncotarget. 2017 Aug 14;8(50):87263-87275. doi: 10.18632/oncotarget.20256. eCollection 2017 Oct 20.
Though there were many researches about the effects of cancer cells on non-small cell lung cancer (NSCLC) currently, it has been rarely reported completed oncogene and its mechanism in tumors by far. Here, we used biological methods with known oncogene of NSCLC to find new oncogene and explore its functionary mechanism in NSCLC.
The study firstly built NSCLC genetic interaction network based on bioinformatics methods and then combined shortest path algorithm with significance test to confirmed core genes that were closely involved with given genes; real-time qPCR was conducted to detect expression levels between patients with NSCLC and normal people; additionally, detection of PARP1's role in migration and invasion was performed by trans-well assays and wound-healing.
Through gene interaction network, it was found that, core genes like PARP1, EGFR and ALK had a direct interaction. TCGA database showed that PARP1 presented strong expression in NSCLC and the expression level of metastatic NSCLC was significantly higher than that of non-metastatic NSCLC. Cell migration of NSCLC in accordance to the scratch test was suppressed by PARP1 silence but stimulated noticeably by PARP1 overexpression. According to Kaplan-meier survival curve, the higher PARP1 expression, the poorer patient survival rate and prognosis. Thus, PARP1 expression had a negative correction with patient survival rate and prognosis.
New oncogene PARP1 was found from known NSCLC oncogene in terms of gene interaction network, demonstrating PARP1's impact on NSCLC cell migration.
目前关于癌细胞对非小细胞肺癌(NSCLC)影响的研究众多,但迄今关于肿瘤中完整致癌基因及其机制的报道仍较为少见。在此,我们运用生物学方法结合已知的NSCLC致癌基因来寻找新的致癌基因,并探究其在NSCLC中的作用机制。
本研究首先基于生物信息学方法构建NSCLC基因相互作用网络,然后将最短路径算法与显著性检验相结合以确定与给定基因密切相关的核心基因;采用实时定量PCR检测NSCLC患者与正常人之间的表达水平;此外,通过Transwell实验和划痕实验检测PARP1在迁移和侵袭中的作用。
通过基因相互作用网络发现,PARP1、EGFR和ALK等核心基因存在直接相互作用。TCGA数据库显示,PARP1在NSCLC中呈高表达,且转移性NSCLC的表达水平显著高于非转移性NSCLC。PARP1沉默可抑制NSCLC细胞在划痕实验中的迁移,而PARP1过表达则显著促进其迁移。根据Kaplan-Meier生存曲线,PARP1表达越高,患者生存率和预后越差。因此,PARP1表达与患者生存率和预后呈负相关。
通过基因相互作用网络从已知的NSCLC致癌基因中发现了新的致癌基因PARP1,证实了PARP1对NSCLC细胞迁移的影响。