中国组织工程研究 ›› 2016, Vol. 20 ›› Issue (7): 993-999.doi: 10.3969/j.issn.2095-4344.2016.07.011

• 组织构建细胞学实验 cytology experiments in tissue construction • 上一篇    下一篇

间充质干细胞条件培养液促进肺癌细胞上皮间质转化

吴佳斌1,王 涛2,杨伟林1,王均洁2,肖婕婓1,王儒琛1,陈振光1   

  1. 1中山大学附属第一医院,广东省广州市 510000;2中山大学干细胞与组织工程研究中心,广东省广州市 510000
  • 收稿日期:2015-01-12 出版日期:2016-02-12 发布日期:2016-02-12
  • 通讯作者: 陈振光,博士,教授,主任医师,硕士生导师,中山大学附属第一医院,广东省广州市 510000
  • 作者简介:吴佳斌,男,河南省焦作市人,汉族,中山大学附属第一医院在读硕士,主要从事肺癌、食管癌转移机制方向的研究。
  • 基金资助:
    国家自然科学基金(青年科学基金项目)(81101774);中国博士后科学基金(2014M552218)

Human bone marrow mesenchymal stem cells promote epithelial mesenchymal  transition in lung cancer cells

Wu Jia-bin1, Wang Tao2, Yang Wei-lin1, Wang Jun-jie2, Xiao Jie-fei1, Wang Ru-chen1, Chen Zhen-guang1   

  1. 1the First Affiliated Hospital of Sun Yat-sen University, Guangzhou 510000, Guangdong Province, China; 2Center for Stem Cell Biology and Tissue Engineering, Sun Yat-sen University, Guangzhou 510000, Guangdong Province, China
  • Received:2015-01-12 Online:2016-02-12 Published:2016-02-12
  • Contact: Chen Zhen-guang, M.D., Professor, Chief physician, Master’s supervisor, the First Affiliated Hospital of Sun Yat-sen University, Guangzhou 510000, Guangdong Province, China
  • About author:Wu Jia-bin, Studying for master’s degree, the First Affiliated Hospital of Sun Yat-sen University, Guangzhou 510000, Guangdong Province, China
  • Supported by:

    the National Natural Science Foundation of China, No. 81101774; the China Postdoctoral Science Foundation, No. 2014M552218

摘要:

文章快速阅读:

文题释义:

骨髓间充质干细胞:存在于骨髓中的非造血类的干细胞,来源于胚胎发育早期中胚层的一类多能干细胞,具有自我更新及多项分化潜能,在不同的刺激条件下可分化为不同的细胞,如成骨细胞、脂肪细胞、软骨细胞,同时表达CD44、CD73、CD54、CD90、CD105而不表达CD45、CD34、CD11b、CD14等造血干细胞标志物。

上皮间质转化:指上皮细胞通过特定程序转化为具有间质表型细胞的生物学过程。在胚胎发育、慢性炎症、组织重建、癌症转移和多种纤维化疾病中发挥了重要作用,其主要的特征有细胞黏附分子E-钙黏素表达降低,间质细胞相关标志物N-钙黏素、波形蛋白等表达上升以及形态上具有间充质细胞的特征等。

 

背景:骨髓间充质干细胞与肿瘤关系的复杂性严重限制了间充质干细胞的应用 ,因此明确间充质干细胞在肿瘤生长与转移中的作用已成为迫切需要解决的问题。
目的:探讨人骨髓来源间充质干细胞对人非小细胞肺癌A549、PAa细胞上皮间质转化的影响。
方法:收集间充质干细胞上清液作为间充质干细胞条件培养液培养肺癌细胞A549和PAa,观察肺癌细胞形态、上皮间质转化标志物E-钙黏素、N-钙黏素、波形蛋白以及其转录因子Slug、Snail、Twist等的表达以及肺癌细胞运动迁移能力的变化。

结果与结论:①与对照组对比:加入条件培养液的肺癌细胞形态发生了间质样变化,且E-钙黏素表达降低,N-钙黏素、波形蛋白mRNA与蛋白表达水平上升,转录因子Slug、Snail、Twist mRNA表达水平也明显增加,同时细胞运动迁移能力增强。②结果表明人骨髓来源间充质干细胞可以促进非小细胞肺癌A549、PAa细胞上皮间质转化过程,增强肺癌细胞运动迁移能力。 

ORCID: 0000-0003-0668-5066(陈振光)

关键词: 组织构建, 组织工程, 间充质干细胞, 肺癌, 上皮间质转化, 转移, A549细胞, PAa细胞, 条件培养, 国家自然科学基金

Abstract:

BACKGROUND: The complex relationship between bone marrow mesenchymal stem cells and cancers severely limit the clinical application of mesenchymal stem cells. So it is urgent to study the role of mesenchymal stem cells in tumor growth and metastasis.
OBJECTIVE: To explore the effect of human bone marrow mesenchymal stem cells on epithelial mesenchymal transition in non-small cell lung cancer A549 and PAa cells.
METHODS: The A549 and PAa cells were cultured with mesenchymal stem cell supernatant (mesenchymal stem cell conditioned medium, MSCs-CM). The cellular morphology was observed under a microscope. The mRNA and protein expression of E-cadherin, N-cadherin, Vimentin, Slug, Snail, and Twist were determined by RT-PCR and western blot. Transwell and wound healing assay were used to detect the change of migration and metastatic ability.
RESULTS AND CONCLUSION: Compared with the control group, the cellular morphology of experimental group showed mesenchymal-like changes. In response to MSCs-CM, there was decreased E-cadherin but increased N-cadherin, Vimentin and Slug, Snail, Twist at mRNA and protein levels compared with the control group (P < 0.05). The migration and metastatic abilities of the experimental group were also increased. So, human bone marrow mesenchymal stem cells can promote epithelial mesenchymal transition in A549 and PAa cells, and enhance the migration and metastatic abilities of A549 and PAa cells.