| 谭力芯,叶艳艳,熊 宇,税桦桦,黄文静,丁大莲,曾宁碧,王 迪.阿霉素通过Wnt/β-catenin信号通路抑制口腔鳞癌干细胞迁移和侵袭[J].,2021,(3):429-435 |
| 阿霉素通过Wnt/β-catenin信号通路抑制口腔鳞癌干细胞迁移和侵袭 |
| Inhibitory Effects of Adriamycin on the Migration and Invasion of Oral Squamous Carcinoma Stem Cells via Wnt/β-catenin Signaling Pathway |
| 投稿时间：2020-05-27 修订日期：2020-06-23 |
| DOI：10.13241/j.cnki.pmb.2021.03.006 |
| 中文关键词: 口腔鳞癌干细胞 阿霉素 Wnt/β-catenin信号通路 迁移 侵袭 |
| 英文关键词: Oral squamous cell carcinoma stem cells Doxorubicin Wnt/ β-catenin signaling pathway Migration Invasion |
| 基金项目:教育部留学回国人员科研启动基金项目(HG2015-002)；院级军科基金项目(SWH2016JSTSYB-40N) |
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| 中文摘要: |
| 摘要 目的：探讨阿霉素对口腔鳞癌干细胞迁移、侵袭、凋亡的影响及其可能的机制。方法：体外培养人口腔鳞癌细胞系SCC25，通过流式细胞术分选CD44-和CD44+细胞，RT-PCR检测CD44-和CD44+细胞的Oct4、CD133、CD44和GAPDH的mRNA表达；检测和比较CD44-和CD44+细胞的克隆形成能力。CD44+细胞用阿霉素或β-catenin抑制剂LF3进行处理，分别使用Transwell和细胞划痕检测细胞侵袭和迁移能力，一步法TUNEL检测细胞凋亡水平，WB检测β-catenin和TCF-4的蛋白表达。结果：流式细胞术成功分离CD44-和CD44+细胞，RT-PCR检测CD44+细胞高表达Oct4、CD133和CD44 mRNA，CD44-细胞弱表达Oct4 mRNA，不表达CD133和CD44 mRNA；CD44+细胞的克隆形成能力显示显著强于CD44-细胞(P<0.05)。阿霉素显著降低了CD44+细胞的侵袭能力和迁移能力(P<0.05)，显著提高了CD44+细胞的凋亡率(P<0.05)；阿霉素显著降低了CD44+细胞β-catenin和TCF-4的蛋白表达 (P<0.05)，LF3对β-catenin和TCF-4蛋白表达的影响与阿霉素比较无显著差异(P>0.05)。结论：阿霉素可能通过抑制Wnt/β-catenin信号通路降低口腔鳞癌干细胞迁移、侵袭能力，促进细胞凋亡。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate the effects of adriamycin on the migration, invasion and apoptosis of oral squamous cell carcinoma stem cells and its related signal pathways. Methods: Human oral squamous cell carcinoma cell line SCC25 was cultured in vitro. CD44- and CD44+ cells were sorted and compared by flow cytometry. RT-PCR was used to detect the mRNA expression levels of Oct4, CD133, CD44, and GAPDH in CD44- and CD44+ cells. CD44- and CD44+ cells were tested for their ability to form clones. CD44+ cells were treated with adriamycin or inhibitor LF3, and Transwell and cell scratches were used to detect cell invasion and migration ability. One-step TUNEL was used to detect apoptosis. WB was used to detect β-catenin and TCF-4 protein expression levels. Results: CD44- and CD44+ cells successfully isolated by flow cytometry. RT-PCR detected that CD44+ cells strongly expressed Oct4, CD133 and CD44 mRNA. CD44- cells weakly expressed Oct4 mRNA, and did not express CD133 and CD44 mRNA. The results of clonal formation analysis showed that CD44+ cells were significantly stronger than CD44- cells (P<0.05). Adriamycin significantly reduced CD44+ cell invasion capacity (P<0.05). Adriamycin significantly reduced CD44+ cell migration capacity (P<0.05). Adriamycin significantly increased CD44+ cell apoptosis levels (P<0.05). Adriamycin significantly reduced β-catenin and TCF-4 protein expression in CD44+ cells (P<0.05), and the expression of β-catenin and TCF-4 protein by LF3 culture was not significantly different with adriamycin (P>0.05). Conclusion: Adriamycin reduces the migration and invasion ability of oral squamous cell carcinoma stem cells by inhibiting the Wnt/ β-catenin signaling pathway, and promotes apoptosis. |
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