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.

参考文献

[1] Johnson DE, Burtness B, Leemans CR, et al.Head and neck squamous cell carcinoma[J]. Nat Rev Dis Primers, 2020, 6(1): 92-140.
[2] Chow LQM.Head and neck cancer[J]. N Engl J Med, 2020, 382(1): 60-72.
[3] 谭晨, 王茹, 房居高. 诱导化疗在局部中晚期头颈鳞状细胞癌治疗中的应用进展[J]. 国际耳鼻咽喉头颈外科杂志, 2019, 43(3): 159-162.
Tan C, Wang R, Fang JG.Progress of the therapeutical effect with induction chemotherapy on advanced head and neck squamous cell carcinoma[J]. International Journal of Otolaryngology-Head and Neck Surgery, 2019, 43(3): 159-162.
[4] Posner MR, Colevas AD, Tishler RB.The role of induction chemotherapy in the curative treatment of squamous cell cancer of the head and neck[J]. Semin Oncol, 2000, 27(4 Suppl 8): 13-24.
[5] Geoffrois L, Martin L, De Raucourt D, et al.Induction chemotherapy followed by cetuximab radiotherapy is not superior to concurrent chemoradiotherapy for head and neck carcinomas: results of the GORTEC 2007-02 phase Ⅲ randomized trial[J]. J Clin Oncol, 2018, 36(31): 3077-3083.
[6] Hitt R, Grau JJ, López-pousa A, et al. A randomized phase Ⅲ trial comparing induction chemotherapy followed by chemoradiotherapy versus chemoradiotherapy alone as treatment of unresectable head and neck cancer[J]. Ann Oncol, 2014, 25(1): 216-225.
[7] Cohen EE, Karrison TG, Kocherginsky M, et al.Phase Ⅲ randomized trial of induction chemotherapy in patients with N2 or N3 locally advanced head and neck cancer[J]. J Clin Oncol, 2014, 32(25): 2735-2743.
[8] Haddad R, O'neill A, Rabinowits G, et al. Induction chemotherapy followed by concurrent chemoradiotherapy (sequential chemoradiotherapy) versus concurrent chemoradiotherapy alone in locally advanced head and neck cancer (PARADIGM): a randomised phase 3 trial[J]. Lancet Oncol, 2013, 14(3): 257-264.
[9] Lefebvre J L, Andry G, Chevalier D, et al.Laryngeal preservation with induction chemotherapy for hypopharyngeal squamous cell carcinoma: 10-year results of EORTC trial 24891[J]. Ann Oncol, 2012, 23(10): 2708-2714.
[10] Posner MR, Hershock DM, Blajman CR, et al.Cisplatin and fluorouracil alone or with docetaxel in head and neck cancer[J]. N Engl J Med, 2007, 357(17): 1705-1715.
[11] Adkins D, Ley J, Neupane P, et al.Palbociclib and cetuximab in platinum-resistant and in cetuximab-resistant human papillomavirus-unrelated head and neck cancer: a multicentre, multigroup, phase 2 trial[J]. Lancet Oncol, 2019, 20(9): 1295-1305.
[12] Posner MR, Lefebvre JL.Docetaxel induction therapy in locally advanced squamous cell carcinoma of the head and neck[J]. Br J Cancer, 2003, 88(1): 11-17.
[13] 董娜, 邵峰. 细胞焦亡的机制和功能[J]. 中国科学(生命科学), 2019, 49(12): 1606-1634.
Dong N, Shao F.Molecular mechanism and immunological function of pyroptosis[J]. Science in China: Life Sciences,2019, 49(12): 1606-1634.
[14] Zhang Z, Zhang Y, Xia S, et al.Gasdermin E suppresses tumour growth by activating anti-tumour immunity[J]. Nature, 2020, 579(7799): 415-420.
[15] Wang Y, Gao W, Shi X, et al.Chemotherapy drugs induce pyroptosis through caspase-3 cleavage of a gasdermin[J]. Nature, 2017, 547(7661): 99-103.
[16] Rogers C, Fernandes-Alnemri T, Mayes L, et al.Cleavage of DFNA5 by caspase-3 during apoptosis mediates progression to secondary necrotic/pyroptotic cell death[J]. Nat Commun, 2017, 8: 14128.
[17] Westbom C, Thompson JK, Leggett A, et al.Inflammasome modulation by chemotherapeutics in malignant mesothelioma[J]. PLoS One, 2015, 10(12): e0145404.
[18] Hu L, Chen M, Chen X, et al.Chemotherapy-induced pyroptosis is mediated by BAK/BAX-caspase-3-GSDME pathway and inhibited by 2-bromopalmitate[J]. Cell Death Dis, 2020, 11(4): 281-297.
[19] Wang L, Li K, Lin X, et al.Metformin induces human esophageal carcinoma cell pyroptosis by targeting the miR-497/PELP1 axis[J]. Cancer Lett, 2019, 450: 22-31.
[20] Johnson DC, Taabazuing CY, Okondo MC, et al.DPP8/DPP9 inhibitor-induced pyroptosis for treatment of acute myeloid leukemia[J]. Nat Med, 2018, 24(8): 1151-1156.
[21] Cai J, Yi M, Tan Y, et al.Natural product triptolide induces GSDME-mediated pyroptosis in head and neck cancer through suppressing mitochondrial hexokinase-Ⅱ[J]. J Exp Clin Cancer Res, 2021, 40(1): 190-206.
[22] Zhang CC, Li CG, Wang YF, et al.Chemotherapeutic paclitaxel and cisplatin differentially induce pyroptosis in A549 lung cancer cells via caspase-3/GSDME activation[J]. Apoptosis, 2019, 24(3-4): 312-325.
[23] Yu P, Wang HY, Tian M, et al.Eukaryotic elongation factor-2 kinase regulates the cross-talk between autophagy and pyroptosis in doxorubicin-treated human melanoma cells in vitro[J]. Acta Pharmacol Sin, 2019, 40(9): 1237-1244.
[24] Jiang M, Qi L, Li L, et al.The caspase-3/GSDME signal pathway as a switch between apoptosis and pyroptosis in cancer[J]. Cell Death Discov, 2020, 6: 112-123.
[25] 邢栋, 王焕焕, 吕勃. 细胞程序性坏死与细胞焦亡[J]. 生理科学进展, 2020, 51(2): 113-116.
Xing D, Wang HH, Lyu B.Necroptosis and pyroptosis[J]. Progress in Physiological Sciences, 2020, 51(2): 113-116.
[26] Kovacs SB, Miao EA.Gasdermins: effectors of pyroptosis[J]. Trends Cell Biol, 2017, 27(9): 673-684.
[27] Yu J, Li S, Qi J, et al.Cleavage of GSDME by caspase-3 determines lobaplatin-induced pyroptosis in colon cancer cells[J]. Cell Death Dis, 2019, 10(3): 193-213.
[28] Bray F, Ferlay J, Soerjomataram I, et al.Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries[J]. CA Cancer J Clin, 2018, 68(6): 394-424.
[29] Nagata S.Apoptosis and clearance of apoptotic cells[J]. Annu Rev Immunol, 2018, 36: 489-517.
[30] Mizumachi T, Suzuki S, Naito A, et al.Increased mitochondrial DNA induces acquired docetaxel resistance in head and neck cancer cells[J]. Oncogene, 2008, 27(6): 831-838.
[31] Zhu X, Yu Z, Feng L, et al.Chitosan-based nanoparticle co-delivery of docetaxel and curcumin ameliorates anti-tumor chemoimmunotherapy in lung cancer[J]. Carbohydr Polym, 2021, 268: 118237.
[32] Shen X, Wang H, Weng C, et al.Caspase 3/GSDME-dependent pyroptosis contributes to chemotherapy drug-induced nephrotoxicity[J]. Cell Death Dis, 2021, 12(2): 186-202.
[33] Erkes DA, Cai W, Sanchez IM, et al.Mutant BRAF and MEK inhibitors regulate the tumor immune microenvironment via pyroptosis[J]. Cancer Discov, 2020, 10(2): 254-269.

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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