多西他赛对头颈鳞癌细胞焦亡的影响及作用机制探讨

doi:10.19438/j.cjoms.2023.02.001

中国口腔颌面外科杂志 ›› 2023, Vol. 21 ›› Issue (2): 105-111.doi: 10.19438/j.cjoms.2023.02.001

多西他赛对头颈鳞癌细胞焦亡的影响及作用机制探讨

高嘉敏, 姚艳丽, 王玉珏, 张志愿, 孙树洋

上海交通大学医学院附属第九人民医院口腔颌面-头颈肿瘤科,上海交通大学口腔医学院, 国家口腔医学中心,国家口腔疾病临床医学研究中心,上海市口腔医学重点实验室, 上海市口腔医学研究所,上海 200011

收稿日期:2022-10-17 修回日期:2022-12-26 出版日期:2023-03-20 发布日期:2023-06-12
通讯作者: 孙树洋,E-mail: sunshuyang@sjtu.edu.cn;张志愿,E-mail: zhzhy0502@163.com。^#共同通信作者
作者简介:高嘉敏（1997-）,女,硕士研究生,E-mail：gaojiamin18916212273@sjtu.edu.cn
基金资助:
国家自然科学基金面上项目（81872199）; 国家自然科学基金重点项目（82030085）; 国家重点研发计划（2017YFC0908500）

Effect of docetaxel on cell pyroptosis of head and neck squamous cell carcinoma and its mechanism

GAO Jia-min, YAO Yan-li, WANG Yu-jue, ZHANG Zhi-yuan, SUN Shu-yang

Department of Oromaxillofacial Head and Neck Oncology, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine; College of Stomatology, Shanghai Jiao Tong University; National Center for Stomatology; National Clinical Research Center for Oral Diseases; Shanghai Key Laboratory of Stomatology; Shanghai Research Institute of Stomatology. Shanghai 200011, China

Received:2022-10-17 Revised:2022-12-26 Online:2023-03-20 Published:2023-06-12

摘要/Abstract

摘要： 目的： 探讨多西他赛对头颈鳞癌细胞焦亡的影响及分子机制。方法： 用对照组（0 μmol/L）和不同浓度多西他赛（0.625、1.25、2.5、5、10 μmol/L）处理头颈鳞癌细胞CAL33及SCC154,采用活细胞工作站监测细胞焦亡形态变化。利用蛋白免疫印迹法（Western blot, WB）检测Gasdermin-E（GSDME）、GSDME-N端、Caspase-3及Caspase-3剪切蛋白的蛋白表达;借助活细胞Caspase-3活性与Annexin V细胞凋亡检测试剂盒检测活细胞中Caspase-3活性;采用活性氧（reactive oxygen species,ROS）检测试剂盒检测细胞内ROS水平。使用ROS抑制剂乙酰半胱氨酸抑制细胞ROS的产生,进一步利用WB检测对照组（0 μmol/L）、2.5 μmol/L多西他赛添加5 mmol/L乙酰半胱氨酸处理组、2.5 μmol/L多西他赛不添加乙酰半胱氨酸处理组及5 mmol/L乙酰半胱氨酸不添加多西他赛组处理组内GSDME及GSDME-N端剪切蛋白的蛋白表达,观察ROS抑制对焦亡通路的影响。采用SAS 9.4软件包对数据进行统计学分析。结果： 与对照组相比,多西他赛诱导CAL33及SCC154产生细胞溶胀崩解,胞膜表面出现气泡样凸起的焦亡表型;促进Caspase-3活性;显著上调GSDME-N端及Caspase-3剪切蛋白的蛋白表达量（P<0.05）;诱导ROS水平增加。乙酰半胱氨酸显著降低多西他赛诱导后头颈鳞癌细胞的GSDME-N端蛋白表达量（P<0.05）。结论： 多西他赛可能激活ROS/Caspase-3/GSDME通路,诱导头颈鳞癌细胞焦亡。

关键词: 多西他赛, 头颈鳞癌, 细胞焦亡, Caspase-3, GSDME

Abstract: PURPOSE: To explore the effect of docetaxel on pyroptosis of head and neck squamous carcinoma and the underlying molecular mechanism. METHODS: Head and neck squamous carcinoma cell lines (CAL33 and SCC154) were treated with control group(0 μmol/L) and different concentrations of docetaxel (0.625, 1.25, 2.5, 5 and 10 μmol/L). The morphological change of cell proptosis was observed by living cell workstation. Western blot was used to assess the protein expression of GSDME, GSDME-N terminal, caspase-3 and cleaved caspase-3. The expression of caspase-3 in living cells was monitored by Caspase-3 Activity and Apoptosis Detection Kit for Live Cell. The level of reactive oxygen species(ROS) was quantified by Reactive Oxygen Species Assay Kit. To determine whether the inhibition of ROS had an effect on pyroptosis induced by docetaxel, acetylcysteine (NAC) was used to inhibit the ROS accumulation and the protein expression of GSDME and GSDME-N terminal of control group(0 μmol/L) was tested, group of docetaxel (2.5 μmol/L) without NAC(5 mmol/L), group of docetaxel with NAC and group of NAC without docetaxel by Western blot. SAS 9.4 software package was used for data analysis. RESULTS: Compared with the control group, docetaxel induced cell pyroptosis of CAL33 and SCC154, which was characterized by membrane rupture and surface protrusions. Docetaxel also promoted caspase-3 activity in the living cells, while the GSDME-N terminal and Cleaved-caspase-3 were upregulated(P<0.05). The level of ROS was increased by docetaxel, and high expression of GSDME-N terminal was significantly reversed by acetylcysteine(P<0.05). CONCLUSIONS: Docetaxel may induce pyroptosis of head and neck squamous carcinoma cells by activating ROS/Caspase-3/GSDME pathway.

Key words: Docetaxel, Head and neck squamous cell cancer, Cell pyroptosis, Caspase-3, GSDME

中图分类号:

R739.8

高嘉敏, 姚艳丽, 王玉珏, 张志愿, 孙树洋. 多西他赛对头颈鳞癌细胞焦亡的影响及作用机制探讨[J]. 中国口腔颌面外科杂志, 2023, 21(2): 105-111.

GAO Jia-min, YAO Yan-li, WANG Yu-jue, ZHANG Zhi-yuan, SUN Shu-yang. Effect of docetaxel on cell pyroptosis of head and neck squamous cell carcinoma and its mechanism[J]. China J Oral Maxillofac Surg, 2023, 21(2): 105-111.

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多西他赛对头颈鳞癌细胞焦亡的影响及作用机制探讨

Effect of docetaxel on cell pyroptosis of head and neck squamous cell carcinoma and its mechanism

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