多参数流式细胞术分析肺癌幼稚细胞免疫表型CD133、CD34、CD44的表达

Multiparametric Flow Cytometry Analyzes the Expressions of Immunophenotype CD133,CD34,CD44 in Lung Cancer Naive Cells

目的研究CD133、CD34、CD44在肺癌组织中的表达及其不同表达组合所构成的细胞亚群。方法收集术前病理确诊的肺癌新鲜组织40例,外周正常肺组织10例,制备成单细胞悬液,选用CD133、CD34、CD44、CD117、CD43、CD45、LIN(CD2,CD3,CD31,CD64)单克隆抗体荧光染色,采用流式细胞仪分析上述表面抗原在肺癌组织及正常肺组织细胞中的分布,通过流式双参数图分析CD133、CD34、CD44双抗原组合表达形成的细胞亚群。结果肺癌实质细胞中(LIN系列及CD45阴性),CD133、CD34、CD44表达弱阳性,CD117、CD43阴性;非参数秩和检验示,CD133、CD34、CD44在肺癌及正常肺组织幼稚细胞中表达水平差异无统计学意义;在肺鳞癌与肺腺癌细胞中,CD44+、CD133-CD34-、CD133+CD44+、CD133+CD44-、CD133+CD34+、CD133-CD34+及CD133-CD44+的表达量不同(P<0.05),其余表型的差异均无统计学意义。聚类分析显示,肺癌幼稚细胞可主要归类为4种亚型。结论CD133、CD34、CD44可能是肺癌幼稚细胞标记,有利于更好研究肺癌细胞发育和演变的过程。

Objective To study the expression of CD133, CD34 and CD44 in lung cancer, and distribution of cell subpopulations that generated by combination of different markers. Methods By collecting 10 normal lung tissues and 40 lung cancer fresh samples that were diagnosed pathologically, preparing single cell suspension, and labelling cells with CD133, CD34, CD44, CDl17, CD43, CD45 and LIN（CD2, CD3, CD31, CD64） monoclone antibodies, we analyzed the distribution of the above-mentioned antibodies in lung cancer cells on FCM. By double-parameters plots, we analyzed the naive cell subpopulations that were generated through combination of positive and negative antigen expression. Results In lung cancer parenchymal-cells （LIN^-CD45^-） expressed CD133, CD34 and CD44 dim positive and, expressed CDl17 and CD43 negative. The mann-Whitney rank test results shows that there was no significant difference for the expression of CD133, CD34, CD44 between lung cancer and normal lung tissue, and CD44^＋, CD133^-CD34^-, CD133^＋CD44^＋ , CD133^＋CD44^- , CD133^＋CD34^＋ , CD133^-CD34^＋ and CD133^-CD44^＋ expressed differently （P 〈 0. 05） between lung adenoearclnoma and lung squamous cell carcinoma. The cluster analysis results suggest that they are classified to 4 subsets. Conclusion CD133, CD34, CD44 could be the marker of lung cancer naive cells, which could contribute to study lung cancer cell development.