解剖学报

解剖学报 ›› 2021, Vol. 52 ›› Issue (2): 237-243.doi: 10.16098/j.issn.0529-1356.2021.02.012

• 肿瘤生物学 • 上一篇    下一篇

轴丝动力蛋白中链基因1在肺腺癌中低表达且抑制肺腺癌细胞侵袭能力

张琳1 王雪婷1 王晓栋1 王玉1 李春涛1 尹崇高2 李洪利3*   

  1. 1.潍坊医学院生命科学与技术学院2017级生物制药专业; 2.护理学院; 3.医学研究实验中心，山东 潍坊 261053
  • 收稿日期:2020-04-06 修回日期:2020-06-17 出版日期:2021-04-06 发布日期:2021-04-06
  • 通讯作者: 李洪利 E-mail:lihongli@wfmc.edu.cn
  • 基金资助:
    国家自然科学基金;国家自然科学基金;国家自然科学基金;山东省自然科学基金;潍坊市科学技术发展基金;潍坊医学院大学生科技创新基金;潍坊医学院大学生科技创新基金;国家级大学生创新训练计划基金

Lower expression of dynein axonemal intermediate china 1 in lung adenocarcinoma and inhibits the invasion of lung adenocarcinoma cells

ZHANG Lin1  WANG Xue-ting1  WANG Xiao-dong1  WANG Yu1  LI Chun-tao1 YIN Chong-gao2  LI Hong-Li3*   

  1. 1.College of Biological Science and Technology, Bipharmaceutical Class of 2017; 2.College of Nursing; 3.Medical Research Center, Weifang Medical University，Shandong Weifang 261053，China
  • Received:2020-04-06 Revised:2020-06-17 Online:2021-04-06 Published:2021-04-06
  • Contact: LI Hong-Li E-mail:lihongli@wfmc.edu.cn

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摘要：

 目的  探讨轴丝动力蛋白中链基因1（DNAI1）在肺腺癌（LUAD）中的表达情况以及对肺腺癌侵袭能力的影响。   方法  微阵列基因芯片筛选肺腺癌组织(3例)与癌旁组织（3例）的差异表达基因；聚类热图(heatmap)、火山图（volcano plot）展示筛选后mRNA的表达和分布情况；利用DAVID数据库进行基因本体论（GO）和京都基因与基因组百科全书（KEGG）分析； STRING数据库和Cytoscape 3.6.1软件进行蛋白相互作用（PPI）分析并筛选核心基因（Hub基因)；结合DEGs、Ualcan数据库中各Hub基因在肺腺癌中的差异表达情况选择目的基因DNAI1；Real-time PCR、Western blotting检测DNAI1在人正常肺上皮细胞系BEAS-2B和肺腺癌细胞系H1299、A549中的表达情况；过表达DNAI1后观察细胞形态学变化；侵袭实验检测对A549细胞侵袭能力的影响。   结果  基因芯片结果筛选出86个上调基因和396个下调基因；差异基因参与RNA聚合酶Ⅱ启动子转录的正调控，凋亡过程的负调控，蛋白结合等功能，在胞内广泛分布，与代谢途径和癌症相关途径等信号通路有密切联系；DEGs和Ualcan数据库显示，筛选出的10个Hub基因中，DNAI1在肺腺癌组织中下调且差异表达较为显著；Real-time PCR和Western blotting结果显示，相较于人正常肺上皮细胞系BEAS-2B，DNAI1在肺腺癌细胞系H1299、A549中低表达；过表达DNAI1后A549细胞变圆，并有少数脱落；侵袭实验结果表明，过表达DNAI1后A549细胞侵袭能力显著降低。  结论  DNAI1在肺腺癌中低表达且抑制肺腺癌细胞侵袭能力。

关键词: 轴丝动力蛋白中链基因1, 肺腺癌, 基因芯片, 免疫印迹法, 实时定量聚合酶链反应

Abstract:

Objective  To investigate the expression of dynein axonemal intermediate chain 1(DNAI1) in lung adenocarcinoma(LUAD) and its influence on invasive ability of lung adenocarcinoma.   Methods  Microarray gene chip analysis was used to screen different expression genes in lung adenocarcinoma(3 samples) and adjacent normal tissues(3 samples); Heatmap and volcano plot were performed demonstrate the mRNA expression and distribution after screening; DAVID database used for Gene Ontology(GO) and Kyoto Encyclopedia of Genes of Genomes(KEGG) analysis; STRING database and Cytoscape 3.6.1 software for protein-protein interaction (PPI) analysis and screening of Hub genes; Objective genes were selected based on the differential expression of each Hub gene in lung adenocarcinoma in DEGs and Ualcan database ; Real-time PCR and Western blotting were used to detect the expression of DNAI1 in BEAS-2B, H1299 and A549; observe the morphological changes after DNAI1 overexpression; Transwell invasion assay was used to detect the change of invasion ability of A549 cells after DNAI1 overexpression.   Results  The microarray result  showed that there were 86 up-regulated genes and 396 down-regulated genes; different genes were involved in the RNA polymerase Ⅱ promoter positive regulation of transcription, apoptosis process of negative regulation, protein binding, and other functions, widely distributed within the cell, and associated with the metabolic pathway, cancer and other signal pathways were closely related; DEGs database and Ualcan database showed that DNAI1 was the most downregulated among Hub genes in LUAD; the result  of Real-time PCR and Western blotting showed that DNAI1 had lower expression in H1299 and A549 compared with BEAS-2B; after DNAI1 overexpression, A549 cells became round and a few shed off; invasion assay showed that the invasion ability of A549 cells was significantly reduced.    Conclusion DNAI1 has a lower expression and inhibits the ability of invasion in LUAD, and this study can provide a potential molecular target and provide a theoretical basis for targeted therapy of LUAD.

Key words: Dynein axonemal intermediate china 1, Lung adenocarcinoma, Microarray, Western blotting, Real-time PCR

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