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目的:探讨喉癌干细胞的分选方法,分析顺铂、放射线联合应用对喉癌肿瘤干细胞的杀伤效应及机制。方法:应用流式细胞仪荧光活化细胞分选技术检测并分选出喉癌Hep-2细胞系中的CD133+细胞和CD133-细胞,并检测CD133+细胞亚群的干细胞特性;采用CCK-8试剂盒分别检测顺铂、放射线对两组细胞的生长抑制率;流式细胞仪检测不同干预方案对2组细胞凋亡率及细胞周期分布情况的影响。结果:CD133+细胞在喉癌Hep-2细胞系中占(2.43±0.77)%,在细胞增殖、分化及体内成瘤试验中CD133+细胞均表现出肿瘤干细胞特性;不同浓度剂量的顺铂、放射线对Hep-2细胞均有抑制作用,并在一定浓度范围内呈剂量依赖性;顺铂、放射线单独或联合应用时,CD133-细胞凋亡率显著高于CD133+细胞(P<0.01),并产生G0/G1期阻滞。结论:CD133+细胞较CD133-细胞更具有明显的肿瘤干细胞特性,对放化疗具有明显的抵抗作用,对凋亡诱导作用不敏感和细胞周期改变为其机制之一。
OBJECTIVE: To investigate the method for the selection of laryngeal cancer stem cells and to analyze the killing effect and mechanism of cisplatin and radiotherapy on cancer stem cells. Methods: CD133 + cells and CD133- cells in laryngeal carcinoma Hep-2 cell line were detected by fluorescence activated cell sorting (FCM), and the stem cell characteristics of CD133 + cell subpopulation were detected. CCK-8 kit The inhibitory rates of cisplatin and radiation on the growth of the two groups were detected respectively. The effects of different interventions on the apoptosis rate and cell cycle distribution of the two groups were detected by flow cytometry. Results: CD133 + cells in the laryngeal cancer Hep-2 cell line (2.43 ± 0.77)%, CD133 + cells showed tumor stem cell characteristics in cell proliferation, differentiation and in vivo tumorigenicity experiments; different concentrations of cisplatin, Hep-2 cells were inhibited in a dose-dependent manner. When treated with cisplatin or radiation alone or in combination, the apoptosis rate of CD133- cells was significantly higher than that of CD133 + cells (P <0.01), and G0 / G1 arrest. CONCLUSION: CD133 + cells have more obvious characteristics of tumor stem cells than CD133- cells, have obvious resistance to radiotherapy and chemotherapy, are not sensitive to apoptosis induction and cell cycle change is one of the mechanisms.