BACKGROUND The epidermal growth factor receptor (EFGR) mutation was closely related to the invasion and metastasis of lung adenocarcinoma and the biological axis of CXCR4/CXCL12 (chemokine receptor 4/chemokine ligand 12) played an important role in the organ-specific metastasis of the tumor. It was a question surrounding whether there is interaction between them in the process of lung adenocarcinoma metastasis. To investigate the potential molecular mechanisms of EGFR over-expression and EFGR-mutations effects on cell proliferation, migration and invasion, we constructed EGFR over-expression and three EFGR-mutant human lung adenocarcinoma H1299 cell sublines. METHODS EGFR over-expression and three EFGR-mutant (EGFR-E746-A750del, EGFR-T790M and EGFR-L858R) plasmid were designed and transfected H1299 cells with Lipofectamine 2000. H1299 cells transfected with empty vector were negative control (NC), and H1299 cells without transfection were set as blank control (BC). The effects of EGFR over-expression and mutations on the proliferation, migration and invasion of H1299 cells were detected by cell cloning assay, wound healing assay and Transwell assay. The mRNA and protein expression levels of MMP-2, MMP-9, CXCR4 and CXCL12 were detected by RT-PCR and Western blot. RESULTS Compared with negative control group and blank control group, EGFR over-expression and EGFR-E746-A750 deletion have significantly higher colony formation (28±2, 28.33±4.16; respectively) (P<0.05) and the cell migration and invasion ability were significantly increased (P<0.05). RT-PCR and Western blot assay showed that the mRNA and protein expression of MMP-2, MMP-9, CXCR4 and CXCL12 in EGFR over-expression and EGFR-E746-A750 deletion group were remarkably higher than that in negative control and blank control group (P<0.05). CONCLUSIONS EGFR over-expression and 19 exon deletion can promote the expression of MMP-2 and MMP-9 by up-regulating CXCR4/CXCL12 signaling pathway, leading to the change of tumor biological characteristics such as higher proliferation, migration and invasion ability. 【中文题目：肺腺癌细胞EGFR基因过表达和突变
通过介导CXCR4/CXCL12信号通路表达
导致肿瘤生物学行为改变的机制研究】 【中文摘要：背景与目的 表皮生长因子受体（epidermal growth factor receptor, EFGR）突变与肺腺癌侵袭转移密切相关，CXCR4/CXCL12（chemokine receptor 4/chemokine ligand 12）生物学轴在肿瘤器官特异性转移中发挥重要作用，二者在肺腺癌转移过程中是否存在相互作用尚未明确。本研究旨在探索EGFR过表达和不同位点突变对肿瘤细胞增殖、迁移和侵袭等生物学行为的影响，并探讨其潜在机制。方法 构建EGFR过表达、EGFR-E746-A750缺失型（DEL，19号外显子突变）、EGFR-T790M突变型（790M，20号外显子突变）、EGFR-L858R的突变型（LR，21号外显子突变）及EGFR空载质粒，应用Lipofectamine 2000转染H1299肺腺癌细胞。应用细胞克隆实验、细胞划痕实验和Transwell实验分别检测EGFR过表达和突变对H1299细胞增殖、迁移和侵袭能力的影响，并应用RT-PCR和Western blot检测CXCR4、CXCL12，以及该信号通路下游基因MMP-2和MMP-9 mRNA和蛋白表达水平。结果 EGFR过表达和EGFR-E746-A750缺失组细胞克隆形成数目分别为28±2、28.33±4.16，且细胞迁移和侵袭能力显著高于阴性对照组与空白对照组（P<0.05）。RT-PCR和Western blot实验显示EGFR过表达和EGFR-E746-A750缺失组CXCR4、CXCL12、MMP-2和MMP-9的mRNA和蛋白表达水平显著高于阴性对照组和空白对照组（P<0.05）。结论 EGFR基因过表达和19号外显子缺失突变可通过上调CXCR4/CXCL12信号通路，促进MMP-2、MMP-9表达，从而引起肺腺癌细胞的肿瘤生物学特性发生改变，并具有更强的增殖、迁移和侵袭能力。】 【中文关键词：EGFR；过表达；突变；生物学行为；肺肿瘤】.

中文翻译：

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[表皮生长因子受体过表达和突变导致通过CXCR4 / CXCL12信号通路人肺腺癌细胞生物学特性改变的机制]。

背景表皮生长因子受体（EFGR）突变与肺腺癌的侵袭和转移密切相关，CXCR4 / CXCL12（趋化因子受体4 /趋化因子配体12）的生物轴在肺癌的器官特异性转移中起重要作用。瘤。这是一个围绕它们在肺腺癌转移过程中是否存在相互作用的问题。为了研究EGFR过表达和EFGR突变对细胞增殖，迁移和侵袭的潜在分子机制，我们构建了EGFR过表达和三种EFGR突变的人肺腺癌H1299细胞亚系。方法设计EGFR过表达和三个EFGR突变体（EGFR-E746-A750del，EGFR-T790M和EGFR-L858R）质粒，并用Lipofectamine 2000转染H1299细胞。空载体转染的H1299细胞为阴性对照（NC），未转染的H1299细胞设为空白对照（BC）。通过细胞克隆法，伤口愈合法和Transwell法检测EGFR过表达和突变对H1299细胞增殖，迁移和侵袭的影响。用RT-PCR和Western blot检测MMP-2，MMP-9，CXCR4和CXCL12的mRNA和蛋白表达水平。结果与阴性对照组和空白对照组相比，EGFR过表达和EGFR-E746-A750缺失具有明显更高的菌落形成（分别为28±2、28.33±4.16）（P <0.05）以及细胞迁移和侵袭能力显着增加（P <0.05）。RT-PCR和Western blot分析表明，MMP-2，MMP-9，EGFR过表达组和EGFR-E746-A750缺失组的CXCR4和CXCL12明显高于阴性对照组和空白对照组（P <0.05）。结论EGFR的高表达和19个外显子的缺失可以通过上调CXCR4 / CXCL12信号通路来促进MMP-2和MMP-9的表达，从而导致肿瘤生物学特性的改变，如更高的增殖，迁移和侵袭能力。【中文摘要：肺腺癌EGFR基因过表达和突变通过介导CXCR4 / CXCL12信号通路表达导致肿瘤生物学行为改变的机制研究】【中文摘要：背景与目的表皮生长因子受体（表皮生长因子受体） ，