喉癌中侧群细胞的分选及其体外生物学特性

发布时间：2018-04-20 05:16

本文选题：喉肿瘤 + 流式分选术　；参考：《临床耳鼻咽喉头颈外科杂志》2014年18期

【摘要】：目的:探讨喉癌细胞株Hep-2中干细胞样侧群(SP)细胞的分选方法及其干细胞特性。方法:制备喉癌Hep-2单细胞悬液,经Hoechst 33342染色后,用荧光激活细胞分选技术分选出人喉癌SP和非侧群(NSP)细胞,测定SP细胞亚群所占的比例。电子显微镜下观察SP及NSP细胞形态,检测两群细胞的细胞周期、CCK8检测细胞的增殖能力。荧光定量PCR检测SP及NSP细胞中的ABCG2mRNA的相对量,结果用2-ΔΔCt计算。结果:流式细胞仪分析结果表明,SP细胞在Hep-2细胞株中占(13.17±1.85)%。生长曲线(CCK8法)示,SP细胞较NSP细胞有更强的增殖能力;两者所处的细胞周期相似,S期细胞比例及增殖指数两者相比,t分别为0.678、0.614(P0.05)。荧光定量PCR示SP细胞中ABCG2mRNA含量高于NSP细胞。结论:人Hep-2细胞株中存在一部分能将Hoechst33342排出细胞外的SP细胞。SP细胞较NSP有更强的增殖及体内成瘤能力,具有干细胞活性。SP细胞中ABCG2mRNA表达量明显高于NSP细胞。
[Abstract]:Aim: to investigate the sorting method and stem cell characteristics of stem cell like side group (SPN) cells in laryngeal carcinoma cell line Hep-2. Methods: the single cell suspension of Hep-2 was prepared. After Hoechst 33342 staining, SP and non-SSP cells of laryngeal carcinoma were separated by fluorescence activated cell sorting technique, and the percentage of SP cell subsets was measured. The morphology of SP and NSP cells were observed under electron microscope, and the cell cycle and the proliferative ability of the two groups of cells were detected by CCK8. The relative amount of ABCG2mRNA in SP and NSP cells was detected by fluorescence quantitative PCR. The results were calculated with 2- 螖螖 Ct. Results: the results of flow cytometry showed that SP cells accounted for 13.17 卤1.85% of Hep-2 cell lines. Growth curve (CCK8) showed that SP cells had stronger proliferative ability than NSP cells, and the cell ratio and proliferation index of the two cells were similar to those of NSP cells. Fluorescence quantitative PCR showed that the content of ABCG2mRNA in SP cells was higher than that in NSP cells. Conclusion: there are some SP cells in human Hep-2 cell line which can expel Hoechst33342 from the cells. Sp cells have stronger proliferation and tumorigenic ability than NSP. The expression of ABCG2mRNA in the stem cell activity. Sp cells is significantly higher than that in NSP cells.
【作者单位】：广州市第十二人民医院耳鼻咽喉科;
【分类号】：R739.65

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