Rasip1 is a RUNX1 target gene and promotes migration of NSCLC cells.
Rasip1 is a RUNX1 target gene and promotes migration of NSCLC cells.
复制标题
Rasip1 是 RUNX1 靶基因，促进 NSCLC 细胞的迁移。
DOI：
10.2147/cmar.s168438
复制
发表时间：
2018
影响因子：
3.3
通讯作者：
Gu L
中科院分区：
文献类型：
--
作者：
Chen Y;Zhang L;Liu L;Sun S;Zhao X;Wang Y;Zhang Y;Du J;Gu L
Runt 相关转录因子 1 (RUNX1) 是造血的重要调节因子，在非小细胞肺癌 (NSCLC) 患者中过度表达，并且与增强的转移能力相关。 Ras相互作用蛋白1（Rasip1）是一种潜在的癌基因，是血管形成所必需的，最近研究表明Rasip1在NSCLC患者中广泛表达。我们注意到 Rasip1 启动子包含几个潜在的 RUNX1 结合序列。然而，Rasip1和RUNX1在NSCLC中的关系仍不清楚。本研究探讨了RUNX1参与Rasip1表达的潜在功能以及Rasip1在肺癌细胞中的潜在作用。通过定量逆转录聚合酶链反应 (qRT-PCR) 和蛋白质印迹分析 NSCLC 细胞系中的 Rasip1 和 RUNX1 表达。用质粒或干扰RNA (siRNA)转染A549和H1299细胞以上调或下调Rasip1和RUNX1的表达。通过transwell和伤口愈合测定评估细胞运动性。通过免疫荧光检测 Rasip1 和 RUNX1 的位置。同时，使用抗 RUNX1 抗体进行染色质免疫沉淀。构建Rasip1启动子，裂解细胞用于分析荧光素酶活性。在这项研究中，我们发现 RUNX1 的异位表达或敲低分别导致 Rasip1 表达的显着增加或减少。 RUNX1 直接与 Rasip1 启动子内的特定 DNA 序列结合并调节其转录。此外，Rasip1 的沉默通过 Rac1 通路的失活抑制了 RUNX1 过表达的 NSCLC 细胞的迁移。此外，我们发现 Rasip1 在 NSCLC 细胞系中普遍表达并增强细胞迁移。此外，EGFR 信号传导参与 Rasip1 的表达和亚细胞定位。我们的数据表明 Rasip1 部分受转录因子 RUNX1 调节，可能被开发为 NSCLC 的治疗靶点。
Runt-related transcription factor 1 (RUNX1), an essential regulator of hematopoiesis, is overexpressed in patients with nonsmall-cell lung cancer (NSCLC) and is correlated with enhanced metastatic ability. Ras-interacting protein 1 (Rasip1), a potential oncogene, is required for blood vessel formation, and recently, it has been shown that Rasip1 is widely expressed in NSCLC patients. We noticed that Rasip1 promoter contains several potential RUNX1-binding sequences. However, the relationship between Rasip1 and RUNX1 in NSCLC is still unknown. In this study, the potential function of RUNX1 involving in Rasip1 expression and the potential role of Rasip1 in lung cancer cells were investigated. Rasip1 and RUNX1 expressions were analyzed by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and Western blotting in NSCLC cells lines. A549 and H1299 cells were transfected with plasmids or interfering RNA (siRNA) to upregulate or downregulate the expression of Rasip1 and RUNX1. Cell motility was assessed by transwell and wound-healing assay. Location of Rasip1 and RUNX1 was detected via immunofluorescence. Meanwhile, chromatin immunoprecipitation was done using an anti-RUNX1 antibody. Rasip1 promoter was constructed, and cells were lysed for the analysis of luciferase activity. In this study, we showed that ectopic expression or knockdown of RUNX1 resulted in a significant increase or reduction in Rasip1 expression, respectively. RUNX1 bound directly to a specific DNA sequence within Rasip1 promoter and modulated its transcription. Furthermore, silencing of Rasip1 inhibited the migration of RUNX1-overexpressing NSCLC cells through inactivation of Rac1 pathway. Moreover, we found that Rasip1 was expressed ubiquitously in NSCLC cells lines and enhanced cell migration. In addition, EGFR signaling was involved both in the expression and the subcellular localization of Rasip1. Our data indicated that Rasip1 is regulated in part by the transcription factor RUNX1 and might be developed as a therapeutic target for NSCLC.