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目的从小鼠肺腺癌细胞株(lewis lung carcinoma,LLC)中分离并鉴定具有高转移潜能的肿瘤干细胞样细胞亚群。方法流式分析Sca1、CXCR4双阳性细胞比例,激光共聚焦检测小鼠肺癌组织中双阳性细胞表达情况;以CXCR4作为磁珠分选细胞的表面标志,检测分选前后细胞活性,观察分析CXCR4阳性亚群的恶性生物学行为。结果LLC细胞中Sca-1、CXCR4双阳性细胞比例为0.1%,CXCR4阳性细胞为0.18%,小鼠肺癌组织存在荧光双染色细胞;分选前细胞活性为(95.58±0.87)%,分选后活力为(94.96±0.76)%(P>0.05);CXCR4阳性亚群在无血清中呈球状生长,CXCR阴性亚群1×105即不可成瘤,CXCR4阳性亚群在7×103仍可成瘤;CXCR4阴性和阳性亚群分别以5×105、2×104密度各接种3只小鼠,前者无转移,而后者2只发生肺转移,1只发生耳转移。结论Lewis肺癌细胞中的CXCR4阳性亚群具有一定的自我更新和转移能力,具备癌转移性干细胞某些特性。
Objective To isolate and identify tumor stem cell-like subsets with high metastatic potential from mouse lung adenocarcinoma (LLC). Methods Flow cytometry was used to analyze the ratio of Sca1 and CXCR4 double positive cells, and the expression of double positive cells in lung cancer tissues was detected by confocal laser scanning microscope. CXCR4 was used as the surface marker of the magnetic separation cells. The cell viability was detected before and after sorting. CXCR4 positive Subsets of malignant biological behavior. Results The percentage of Sca-1 and CXCR4 double positive cells in LLC cells was 0.1% and that of CXCR4 positive cells was 0.18%. The number of double stained cells in lung cancer tissues was (95.58 ± 0.87)%, The viability was (94.96 ± 0.76)% (P> 0.05). The CXCR4-positive subpopulation was spheroid in serum-free medium. The CXCR negative subgroup 1 × 105 could not form tumor. ; The negative and positive subpopulations of CXCR4 were inoculated with 3 mice at a density of 5 × 10 5 and 2 × 10 4, respectively. The former had no metastasis, while the latter two had lung metastases and one had ear metastases. Conclusion CXCR4 positive subpopulations in Lewis lung cancer cells have certain self-renewal and metastatic potential, and possess some characteristics of cancer metastatic stem cells.