Alantolactone induces concurrent apoptosis and GSDME-dependent pyroptosis of anaplastic thyroid cancer through ROS mitochondria-dependent caspase pathway

doi:10.1016/j.phymed.2022.154528

	Alantolactone induces concurrent apoptosis and GSDME-dependent pyroptosis of anaplastic thyroid cancer through ROS mitochondria-dependent caspase pathway
	Hu, Yiqun 6; Wen, Qingliang 7,8; Cai, Yefeng 6,9; Liu, Yunye 10; Ma, Wenli 2; Li, Qinglin 7,8; Song, Fahuan 3,4,10; Guo, Yawen 3,4,10; Zhu, Lei 1; Ge, Jingyan 5
刊名	PHYTOMEDICINE
	2023
卷号	108
关键词	Alantolactone Anaplastic thyroid cancer ROS Pyroptosis Apoptosis
ISSN号	0944-7113
DOI	10.1016/j.phymed.2022.154528
通讯作者	Ge, Minghua(geminghua@hmc.edu.cn)
英文摘要	Background: Anaplastic thyroid cancer (ATC) is one of the fatal cancers and has not effective treatments. Alantolactone (ATL), a terpenoid extracted from traditional Chinese medicinal herb Inula helenium L., confers significant anti-inflammatory, antibacterial and antitumor activity. However, the activity and mechanisms of ATL in ATC remain unclear. Purpose: To investigate the potential anti-ATC effects in vitro and in vivo and the mechanisms involved. Methods: The anti-proliferative activity of Alantolactone (ATL) against ATC cells was analyzed through CCK-8 and colony formation assays. Flow cytometry assay was performed to assess the cell cycle, cell apoptosis, ROS, and mitochondrial membrane potential (Delta Psi m), whereas the cellular localization of cytochrome c and calreticulin were determined using cellular immunofluorescence assays. The lactate dehydrogenase (LDH) enzyme activity in the cell culture medium was measured using a commercial LDH kit, whereas ELISA was conducted to assess the secretory function of IL-1 beta. Western blot assays were conducted to determine the expression or regulation of proteins associated with apoptosis and pyroptosis. Subcutaneous tumor model of nude mice was established to evaluate the anticancer activity of ATL in vivo. The expression of Ki67, cyclin B1, cleaved-PARP, cleaved-caspase 3, and IL-1 beta in the animal tumor tissues was profiled using immunohistochemistry analyses. Results: Our data showed that ATL significantly inhibited the proliferation and colony formation activity of ATC cells. ATL induced ATC cell cycle arrest at G2/M phase, and downregulated the expression of cyclin B1 and CDC2. Furthermore, ATL induced concurrent apoptosis and pyroptosis in the ATC cells, and the cleavage of PARP and GSDME. It also significantly increased the release of LDH and IL-1 beta. Mechanically, ATL-mediated increase in ROS suppressed the Bcl-2/Bax ratio, downregulated the mitochondrial membrane potential and increased the release of cytochrome c, leading to caspase 9 and caspase 3 cleavage. We also found that ATL induced the translocation of an immunogenic cell death marker (calreticulin) to the cell membrane. In addition, it inhibited the growth of the ATC subcutaneous xenograft model, and activated proteins associated with apoptosis and pyroptosis, with a high safety profile. Conclusion: Taken together, these results firstly demonstrated that ATL exerted an anti-ATC activity by inducing concurrent apoptosis and GSDME-dependent pyroptosis through ROS-mediated mitochondria-dependent caspase activation. Meanwhile, these cell deaths exhibited obvious characteristics of immunogenic cell death, which may synergistically increase the potential of cancer immunotherapy in ATC. Further studies are needed to explore deeper mechanisms for the anti-ATC activity of ATL.
资助项目	National Natural Science Foundation of China[81872170] ; National Natural Science Foundation of China[82103199] ; National Natural Science Foundation of China[21877100] ; National Natural Science Foundation of China - Zhejiang Joint Fund[U20A20382] ; Key Research and Development Program of Zhejiang Province[2021C03081] ; Natural Science Foundation of Zhejiang Province[LQ22H160063] ; Zhejiang Medical and Health Science and Technology Project[2021KY524] ; Zhejiang Medical and Health Science and Technology Project[2022KY064] ; Zhejiang Medical and Health Science and Technology Project[2023RC001] ; Project of Administration of Traditional Chinese Medicine of Zhejiang Province of China[2021ZQ009] ; Project of Administration of Traditional Chinese Medicine of Zhejiang Province of China[2021ZQ011] ; Project of Administration of Traditional Chinese Medicine of Zhejiang Province of China[2023ZL240]
WOS关键词	CELL-DEATH ; MEDIATED APOPTOSIS ; ACTIVATION ; CLEAVAGE
WOS研究方向	Plant Sciences ; Pharmacology & Pharmacy ; Integrative & Complementary Medicine
语种	英语
出版者	ELSEVIER GMBH
WOS记录号	WOS:000883783300001
资助机构	National Natural Science Foundation of China ; National Natural Science Foundation of China - Zhejiang Joint Fund ; Key Research and Development Program of Zhejiang Province ; Natural Science Foundation of Zhejiang Province ; Zhejiang Medical and Health Science and Technology Project ; Project of Administration of Traditional Chinese Medicine of Zhejiang Province of China
内容类型	期刊论文
源URL	[http://ir.hfcas.ac.cn:8080/handle/334002/130349]
专题	中国科学院合肥物质科学研究院
通讯作者	Ge, Minghua
作者单位	1.Wenzhou Med Univ, Hosp 5, Lishui Cent Hosp, Dept Thyroid Surg, Lishui 323000, Zhejiang, Peoples R China 2.Bengbu Med Coll, Bengbu 233030, Anhui, Peoples R China 3.Zhejiang Univ, Sch Med, Dept Publ Hlth, Hangzhou 310014, Peoples R China 4.Key Lab Endocrine Gland Dis Zhejiang Prov, Hangzhou 310014, Peoples R China 5.Zhejiang Univ Technol, Coll Biotechnol & Bioengn, Key Lab Bioorgan Synth Zhejiang Prov, Hangzhou 310014, Peoples R China 6.Zhejiang Chinese Med Univ, Clin Med Coll 2, Hangzhou 310053, Zhejiang, Peoples R China 7.Univ Chinese Acad Sci, Canc Hosp, Zhejiang Canc Hosp, Hangzhou, Peoples R China 8.Chinese Acad Sci, Inst Basic Med & Canc IBMC, Hangzhou 310022, Zhejiang, Peoples R China 9.Wenzhou Med Univ, Affiliated Hosp 1, Dept Thyroid Surg, Wenzhou 325000, Zhejiang, Peoples R China 10.Hangzhou Med Coll, Affiliated Peoples Hosp, Zhejiang Prov Peoples Hosp, Otolaryngol & Head & Neck Ctr,Cancer Ctr,Dept Head, Hangzhou 310014, Zhejiang, Peoples R China
推荐引用方式 GB/T 7714	Hu, Yiqun,Wen, Qingliang,Cai, Yefeng,et al. Alantolactone induces concurrent apoptosis and GSDME-dependent pyroptosis of anaplastic thyroid cancer through ROS mitochondria-dependent caspase pathway[J]. PHYTOMEDICINE,2023,108.
APA	Hu, Yiqun.,Wen, Qingliang.,Cai, Yefeng.,Liu, Yunye.,Ma, Wenli.,...&Ge, Minghua.(2023).Alantolactone induces concurrent apoptosis and GSDME-dependent pyroptosis of anaplastic thyroid cancer through ROS mitochondria-dependent caspase pathway.PHYTOMEDICINE,108.
MLA	Hu, Yiqun,et al."Alantolactone induces concurrent apoptosis and GSDME-dependent pyroptosis of anaplastic thyroid cancer through ROS mitochondria-dependent caspase pathway".PHYTOMEDICINE 108(2023).