Lin Xiangxiao, Liu Xincun, Gong Cunqi
Respiratory Department, Affiliated Hospital of Jining Medical University, Jining, Shandong 272000, P.R. China.
Department of Clinical Laboratory, Jining No. 1 People's Hospital, Jining, Shandong 272000, P.R. China.
Oncol Lett. 2018 Jul;16(1):536-542. doi: 10.3892/ol.2018.8693. Epub 2018 May 10.
The present study aimed to investigate the expression, biological function and mechanism of action of engrailed homeobox 2 (EN2) in non-small cell lung cancer (NSCLC) at the tissue and cellular level. A total of 42 patients who underwent surgical resection of NSCLC tissues between January 2014 and January 2015 were included in the present study. EN2 mRNA expression levels in explanted NSCLC tissues were determined using reverse-transcription quantitative polymerase chain reaction analysis. Adenocarcinoma human alveolar basal epithelial A549 cells were transfected with negative control plasmids or those containing EN2, enabling its overexpression. To assess the effect of EN2 overexpression in A549 cells, a Cell Counting kit-8 assay was used to analyze cellular proliferation, a Transwell assay was used to evaluate cellular migration and invasion and flow cytometry was used to detect the cell cycle distribution. To measure protein expression of EN2 and β-catenin in A549 cells, western blotting was also conducted. EN2 mRNA expression levels in NSCLC tissues were lower than those in normal tissues, and were associated with metastasis, clinical staging and differentiation degrees of NSCLC. Increased expression of EN2 inhibited the proliferation of A549 cells , and suppressed their migration and invasion. Elevated EN2 expression inhibited the proliferation of A549 cells by regulating the G/S phase transition. β-catenin protein expression levels and nuclear translocation in A549 cells were inhibited by EN2 overexpression. The present study demonstrated that expression of EN2 in NSCLC tissues was downregulated and negatively associated with the degree of disease differentiation, lymphatic metastasis and clinical staging. Overexpression of EN2 inhibits the proliferation, migration and invasion of A549 cells, as well as the expression of β-Catenin and nuclear translocation.
本研究旨在从组织和细胞水平探究同源异型盒蛋白2(EN2)在非小细胞肺癌(NSCLC)中的表达、生物学功能及作用机制。本研究纳入了2014年1月至2015年1月期间接受NSCLC组织手术切除的42例患者。采用逆转录定量聚合酶链反应分析测定切除的NSCLC组织中EN2 mRNA表达水平。用阴性对照质粒或含EN2的质粒转染人肺泡基底上皮腺癌A549细胞,使其过表达。为评估EN2过表达对A549细胞的影响,使用细胞计数试剂盒-8法分析细胞增殖,采用Transwell法评估细胞迁移和侵袭,并用流式细胞术检测细胞周期分布。为检测A549细胞中EN2和β-连环蛋白的蛋白表达,还进行了蛋白质印迹分析。NSCLC组织中EN2 mRNA表达水平低于正常组织,且与NSCLC的转移、临床分期及分化程度相关。EN2表达增加抑制了A549细胞的增殖,并抑制其迁移和侵袭。EN2表达升高通过调节G/S期转换抑制A549细胞的增殖。EN2过表达抑制了A549细胞中β-连环蛋白蛋白表达水平及其核转位。本研究表明,EN2在NSCLC组织中的表达下调,且与疾病分化程度、淋巴转移及临床分期呈负相关。EN2过表达抑制A549细胞的增殖、迁移和侵袭,以及β-连环蛋白的表达和核转位。