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论文摘要

桑辛素对喉癌干细胞干性表型调控的作用研究

The Effect of Morusin on Stemness Phenotype of Laryngeal Cancer Stem Cell

作者:陈辰, 许子寒, 王力

Author:CHEN Chen, XU Zi-han, WANG Li

收稿日期:2019-11-22          年卷(期)页码:2020,51(5):650-657

期刊名称:四川大学学报(医学版)

Journal Name:JOURNAL OF SICHUAN UNIVERSITY (MEDICAL SCIENCE EDITION)

关键字:桑辛素, 喉癌, 肿瘤干细胞, 干性表型

Key words:Morusin, Laryngocarcinoma, Cancer stem cell, Stemness phenotype

基金项目:

中文摘要

目的 探索桑辛素对喉癌干细胞干性表型调控的影响。 方法 流式细胞仪分选并检测CD133+喉癌干细胞比例;肿瘤球形成实验检测CD133+喉癌干细胞的自我更新能力;Transwell实验检测CD133+喉癌干细胞的迁移能力;改良MTT实验检测化疗药物对CD133+喉癌干细胞的细胞毒性作用;免疫荧光染色、实时荧光定量PCR(RT-qPCR)及Western blot检测CD133+喉癌干细胞的干细胞标志物表达;在不同浓度的桑辛素处理CD133+喉癌干细胞后(以桑辛素0 μmol/L为对照),通过肿瘤球形成实验、Transwell实验、改良MTT实验及Western blot,分别检测CD133+喉癌干细胞的自我更新能力、迁移能力、化疗药物的细胞毒性作用及干细胞标志物表达变化。 结果 流式细胞仪分选结果显示,CD133+喉癌干细胞占喉癌细胞的比例为(3.50±0.34)%;经过培养富集后,其比例可达(93.20±5.23)%。肿瘤球形成实验结果显示,与喉癌细胞相比,CD133+喉癌干细胞具有增强的自我更新能力 (P<0.001);Transwell实验显示,与喉癌细胞相比,CD133+喉癌干细胞的迁移能力增强(P<0.05);改良MTT实验结果显示,与喉癌细胞相比,CD133+喉癌干细胞抵抗化疗药物(5-氟尿嘧啶及顺铂)的细胞毒性作用(P<0.05);免疫荧光染色、RT-qPCR及Western blot结果显示,干细胞标志物(CD133、ALDH1、Sox2、ABCG2及N-cadherin)在CD133+喉癌干细胞中呈高表达水平。通过不同浓度的桑辛素处理CD133+喉癌干细胞,其自我更新能力降低(P<0.05);迁移能力亦下降(P<0.05);此外,桑辛素处理的CD133+喉癌干细胞对化疗药物的细胞毒性作用更加敏感(P<0.05);Western blot结果显示,不同浓度的桑辛素处理的CD133+喉癌干细胞,其上述的干细胞标志物表达水平下调(P<0.05)。 结论 CD133+喉癌干细胞具有干性表型特征;桑辛素可减弱喉癌干细胞的干性表型,可能与下调其干细胞标志物表达相关。

英文摘要

ObjectiveTo investigate the regulation effect of Morusin on stemness phenotype of laryngeal cancer stem cells.MethodsSeparation and detection the proportion of CD133+laryngeal cancer stem cells through flow cytometry; evaluation the self-renewal ability of CD133+laryngeal cancer stem cells by tumor sphere formation assay; exploring the migration ability of CD133+laryngeal cancer stem cells by Transwell assay; analyzing the cytotoxicity of chemotherapy drugs on CD133+laryngeal cancer stem cells by modified MTT assay; detection of the expression levels of stemness associated markers by immunofluorescence staining, RT-qPCR and Western blot. After treatment with different concentrations of Morusin, cells were performed the above experiments for detection the self-renewal ability, migration ability, cytotoxicity resistance and expression of stemness associated markers.ResultsFlow cytometry analysis showed that the proportion of CD133+laryngeal cancer stem cells was (3.50±0.34)%, while after enrichment, the proportion increased to (93.20±5.23)%. CD133+laryngeal cancer stem cells exhibited better self-renewal ability (P<0 .001) and migratory ability (P<0 .05); they were resistant to the cytotoxicity of chemotherapy drug (P<0 .05), and highly expressed of stemness associated markers. after being treated with morusin, the self-renewal and migratory abilities of cd133+laryngeal cancer stem cells were reduced (P<0 .05). in addition, after treated with morusin, cd133+laryngeal cancer stem cells were more sensitive to chemotherapy drugs; moreover, the expression levels of stemness associated markers were decreased.ConclusionCD133+laryngeal cancer stem cells possessed stemness phenotypic characteristics. Morusin attenuated stemness phenotype of laryngeal cancer stem cells, which may be related to its down-regulation effect on stemness associated markers.

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