摘要
目的 探讨CPNE3对非小细胞肺癌(non-small cell lung cancer, NSCLC)细胞株A549增殖、迁移和侵袭能力的影响及其作用机制。方法 通过癌症和肿瘤基因图谱(the cancer genome atlas, TCGA)的公共数据分析CPNE3在NSCLC中的表达及其与患者预后的关系。利用Western blot方法检测CPNE3在正常支气管上皮细胞株BEAS-2B和肺癌细胞株中的表达。在A549细胞中转染CPNE3的干扰序列,利用CCK-8、克隆形成、Transwell实验检测细胞增殖、迁移和侵袭能力,Western blot法检测下游增殖、转移相关蛋白水平的变化。构建CPNE3过表达稳转细胞株,取10只雌性BALB/c nude小鼠分为对照组和过表达CPNE3组(n=5),构建裸鼠皮下移植瘤模型,观察过表达CPNE3对A549细胞成瘤能力的影响。结果 NSCLC组织中CPNE3表达升高(P<0.05),其高表达与患者的不良预后相关(P<0.01),CPNE3在肺癌细胞株中表达升高(P<0.01),干扰CPNE3后细胞的增殖能力、克隆形成能力下降(P<0.001,P<0.01),迁移和侵袭能力下降(P<0.01,P<0.01),并伴随着p-AKT、p-ERK和p-EGFR蛋白的下调(P<0.001,P<0.001,P<0.01)。过表达CPNE3在体内促进移植瘤的生长(P<0.001),并伴随着p-AKT、p-ERK和p-EGFR蛋白的上调(P<0.05,P<0.05,P<0.01)。结论 干扰CPNE3可以抑制A549细胞的增殖、迁移和侵袭能力,过表达CPNE3可以增强A549细胞在体内的增殖能力,其作用机制与EGFR信号通路及其下游的AKT/ERK信号通路相关。
Objective To investigate the effect of CPNE3 on the proliferation, migration and invasion of non-small cell lung cancer(NSCLC) cell line A549 and its underlying mechanism. Methods The expression of CPNE3 in NSCLC and its relationship with prognosis were analyzed based on the data from the cancer genome atlas(TCGA) database. Western blotting was used to detect the expression of CPNE3 in normal bronchial epithelial cell line BEAS-2 B and lung cancer cell lines. The si-RNAs of CPNE3 were transfected into A549 cells. The abilities of cell proliferation, migration and invasion were detected by CCK-8 assay, colony formation assay, and Transwell assay, respectively, and the expression levels of downstream proteins related to proliferation and metastasis were detected by Western blotting. After the A549 cells with stable overexpression of CPNE3 were established, the cells with overexpression or not were respectively employed to construct lung carcinoma xenograft model in female BALB/c nude mice(n=5 for control and overexpression mice, respectively). Then the tumor growth was observed to determine the effect of CPNE3 overexpression on the tumorigenicity of A549 cells. Results The expression of CPNE3 was increased in NSCLC tissues(P<0.05), and its high expression was correlated with poor prognosis(P<0.01). Consistently, the expression of CPNE3 was increased in lung cancer cell lines(P<0.01). After the knockdown of CPNE3, the proliferation and colony formation(P<0.001, P<0.01), and migration and invasion abilities(P<0.01, P<0.01) were significantly inhibited in A549 cells, accompanied with decreased phosphorylation levels of AKT, ERK, and EGFR(P<0.001, P<0.001, P<0.01). Overexpression of CPNE3 promoted cell proliferation in vivo(P<0.001), and upregulated phosphorylation of AKT, ERK, and EGFR(P<0.05, P<0.05, P<0.01). Conclusion Knocking CPNE3 down can inhibit the proliferation, migration and invasion of A549 cells, while its overexpression can promote the proliferation ability of A549 cells in vivo, which may be related to EGFR signaling pathway and downstream AKT/ERK signaling pathway.
作者
蔡馨
刘泽毅
黄建安
CAI Xin;LIU Zeyi;HUANG Jian’an(Department of Pulmonary and Critical Care Medicine,Institute of Respiratory Diseases,the First Affiliated Hospital of Soochow University,Suzhou,Jiangsu Provmce,215006,China)
出处
《陆军军医大学学报》
CAS
CSCD
北大核心
2022年第24期2507-2513,共7页
Journal of Army Medical University
基金
江苏省卫生健康委科研面上项目(H2019030)
江苏省研究生科研与实践创新计划项目(KYCX22_3220)
关键词
CPNE3
增殖
迁移
侵袭
非小细胞肺癌
CPNE3
proliferation
migration
invasion
lung cancer
non-small cell lung cancer
作者简介
通信作者:黄建安,E-mail:huang_jian_an@163.com
