高/低侵袭转移性肺癌干细胞模型的建立

Establishing High/Low Invasion Lung Tumor Stem Cells model

目的以人肺癌NCI-H446细胞为研究对象,建立一种高/低侵袭性肺癌干细胞(high/low invasion lung tumor stemcells,H/L-ILTSCs)模型,并鉴定其生物学特性。方法首先,利用transwell小室迁移实验分离高/低迁移性NCI-H446细胞。取高/低迁移性NCI-H446细胞采用transwell小室侵袭实验分离高/低侵袭转移性NCI-H446细胞。然后,以CD133、CD44为肺癌干细胞(lung tumor stem cells,LTSCs)表面标志,采用磁珠分选法分离NCI-H446细胞H/L-ILTSCs并进行纯度鉴定。最后,采用MTT法、transwell侵袭及免疫缺陷动物成瘤等实验,检测H-ILTSCs、L-ILTSCs及NCI-H446细胞的生长增殖,体外侵袭转移及体内成瘤能力。结果 H-ILTSCs生长增殖,体外侵袭转移及体内成瘤能力均高于L-ILTSCs及NCI-H446细胞,而L-ILTSCs与NCI-H446无显著差别。结论人肺癌细胞NCI-H446中可以分离和富集H/L-ILTSCs,且二者存在着干性差别。

Objective To establish high/low invasion lung tumor stem cells （H/L - ILTSCs） model and to identify their biological propelties with human lung cancer cell line NCI - H446. Methods First, transwell immigration assay was used to separate high/low im- migration NCI - H446 cells. Then the high/low invasion NCI - H446 ceils were isolated from high/low immigration NCI - H446 cells by transwell invasion assay. Second, we used CD133 and CD44 as markers of human lung cancer stem cells （LTSCs） , separated H/L - ILTSCs by magnetic activated cell sorting and purity identification. Third, the growth and proliferation, tumor invasion in vitro and tumori- genicity potential and metastasis in viva were detected by MTT assay, transwell invasion assay, nude mouse transplanted tumor. Results The growth and proliferation, tumor invasion in vitro and tumorigenicity potential and metastasis in vivo of H - ILTSCs were higher than L - ILTSCs and normal NCI - H446 cells. But it had no difference between L - ILTSCs and normal NCI - H446 ceils. Conclusion H/L - ILTSCs could be isolated and expanded from human lung cancer cell line NCI - H446, and the both cells had different biological prop- erties.