喉癌干细胞在放化疗抵抗中的作用机制

韩瑞; 皇甫辉; 高伟; 张春明; 靳阳子; 李卓; 王斌全

doi:10.13201/j.issn.1001-1781.2014.06.012

喉癌干细胞在放化疗抵抗中的作用机制

- 山西医科大学第一医院耳鼻咽喉-头颈外科山西医科大学耳鼻咽喉研究所(太原, 030001)
基金项目:
山西省自然科学基金 (No:2010011053-1)

详细信息

通讯作者: 皇甫辉, E-mail:13934518228@163.com

中图分类号: R739.65

收稿日期: 2013-10-10

Neoplasms stem cells play an important role in resistance of laryngeal squamous cancer to chemoradiotherapy

- Department of Otorhinolaryngology-Head and Neck Surgery, the First Affiliated Hospital of Shanxi Medical University, Institute of Otolaryngology Affiliated with Shanxi Medical University, Taiyuan, 030001, China

More Information

Corresponding author: HUANGFU Hui, E-mail: 13934518228@163.com

Received Date: 10 October 2013

摘要

摘要: 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。
- 肿瘤干细胞 /
- 喉肿瘤 /
- 联合放化疗 /
- 细胞凋亡 /
- 协同作用
Abstract: Objective:To determine an approach enriching cancer stem cells from laryngeal cancer cell line. To investigate whether laryngeal cancer stem cells in chemoradiotherapy have the characteristic of resistance.Method:CD133⁺ cells and CD133^- cells was detected and isolated from Hep-2 cell line by fluorescence activated cell sorting technology. The cytotoxicities of cisplatin and radiation were investigated by cell counting kit-8(CCK-8) assay. The apoptosis and cell cycle was analyzed with flow cytometry.Result:CD133⁺ cells accounted for a fraction of(2.43±0.77)% in Hep-2 cell line. CD133⁺ cells have a more obvious characteristics of cancer stem cells.Different cisplatin and radiation concentrations of for two cell have inhibition, in a certain concentration range and the dosage dependence. Cisplatin and radiation had synergistic inhibitory effects with CD133^- cells on the growth of two cell. Moreover, cell cycle arrest at G₀/G₁ phase and more apoptosis was induced by synergistic combination. Different concentrations of cetuximab for Hep-2 cells have inhibition, in a certain concentration range and time and the dosage dependence. The half maxial inhibitory concentration (IC₅₀) of cetuximab to Hep-2 cells on 24 h was 1036.84 μg/L.Cisplatin and radiation had synergistic inhibitory effects with cetuximab on the growth of Hep-2 cell line. Moreover, cell cycle arrest at G₀/G₁ phase and more apoptosis was induced by synergistic combination.Conclusion:Compared with CD133^- cells, CD133⁺ cells subpopulation exhibited extraordinary cancer stem.
- neoplastic stem cells /
- laryngeal neoplasms /
- combined radiation and chemotherapy /
- apoptosis /
- synergism

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