肺腺癌细胞三维培养对HOXC10基因表达水平的影响

Three dimensional culture of lung adenocarcinoma cells and its effect on the expression of HOXC10 gene

目的建立肺腺癌A549细胞株三维培养模型,观察和比较二维和三维不同培养条件下肺腺癌细胞的形态、增殖以及HOXC10表达水平的变化。方法二维和三维培养肺腺癌细胞,采用倒置显微镜观察肺腺癌细胞的生长情况,通过CCK8实验检测细胞增殖情况,Real time-PCR和Western blot方法检测HOXC10 mRNA和蛋白表达水平。结果二维单层培养的肺腺癌A549细胞贴壁生长,癌细胞呈梭形。三维细胞培养后,肺腺癌细胞聚集成紧密的多肿瘤细胞微球。HOXC10基因mRNA和蛋白质表达水平在A549细胞三维培养显著高于二维培养。结论HOXC10基因表达在肺腺癌A549细胞三维培养体系显著升高,可能与肿瘤微环境的作用密切相关。

Objective To establish three-dimensional(3D)culture model of adenocarcinoma A549 cells,observe and compare the cellular morphology and proliferation and the expression of HOXC10 gene.Methods The morphology and distribution of lung adenocarcinoma cells in three dimensional culture were identified by inverted microscope.The cell proliferation was detected by CCK8 test.Real time-PCR and western blot was used to assess the expression of HOXC10.Results The lung adenocarcinoma A549 cells in two-dimensional monolayer culture adhered to the wall,and the cancer cells were spindle shaped.After three-dimensional cell culture,lung adenocarcinoma cells gathered into multiple tumor cell microspheres to form hollow alveolar cysts or acinar like structures.The number of cells was increased significantly,and the expression of HOXC10 gene was increased significantly.Conclusion HOXC10 gene expression was significantly increased in the three-dimensional culture system of lung adenocarcinoma A549 cells,which may be closely related to the role of tumor microenvironment.