Cordyceps militaris extract induces apoptosis and pyroptosis via caspase-3/PARP/GSDME pathways in A549 cell line.
Cordyceps militaris extract induces apoptosis and pyroptosis via caspase-3/PARP/GSDME pathways in A549 cell line.
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DOI：
10.1002/fsn3.2636
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发表时间：
2022-01
影响因子：
3.9
通讯作者：
Cai D
中科院分区：
文献类型：
--
作者：
Hu Z;Lai Y;Ma C;Zuo L;Xiao G;Gao H;Xie B;Huang X;Gan H;Huang D;Yao N;Feng B;Ru J;Chen Y;Cai D
蛹虫草（Cordyceps militaris，CM）是我国传统上用于肺癌患者食疗的药物。CM提取物（CME）是CM的水溶性部分，并且被广泛研究。Caspase-3参与的细胞死亡被认为是其主要的抗癌机制，但尚无定论。因此，我们探索其依赖caspase-3的程序性细胞死亡性质（凋亡和焦亡），并在功能丧失实验中验证其依赖caspase-3的性质。通过高效液相色谱-四极杆飞行时间质谱（HPLC-qTOF）检测CME的组分谱。结果表明，CME引起A549细胞出现以细胞凋亡为特征的细胞鼓泡和细胞溶解，并抑制细胞增殖。CME诱导染色质凝聚，使PI+/annexin V+染色在鼓泡细胞中，表明CME引起遗传毒性、细胞凋亡和细胞凋亡。高浓度的CME（200 μg/ml）可使G2/M和G 0期细胞周期阻滞和抑制P53下游增殖蛋白，包括P53、P21、CDC 25 B、CyclinB 1、Bcl-2和BCL 2相关的细胞死亡激动剂（BAD），但1-100 μg/ml的CME对上述蛋白的影响较小。相应地，caspase-3活性和caspase-3下游蛋白，包括pyroptotic effector gasdermin-E（GSDME）和凋亡标志物裂解的聚ADP-核糖聚合酶（PARP），都被CME显著促进。此外，关于在热变性细胞中的膜孔形成，通过CME处理，膜GSDME（与PE-Cy 7缀合的GSDME抗体，用于在流式细胞术中检测）的表达显著增加。相比之下，CME治疗后其他焦亡相关蛋白如P2 X7、NLRP 3、GSDMD和Caspase-1不受影响。此外，TET 2被CME意外地提高。在存在胱天蛋白酶-3抑制剂Ac-DEVD-CHO（Ac-DC）的情况下，CME诱导的细胞毒性、细胞起泡和遗传毒性降低，CME诱导的细胞凋亡（裂解的PARP-1）和细胞凋亡（GSDME-NT）蛋白上调逆转。最后，在HPLC-qTOF实验中鉴定出22种成分，根据之前的参考文献将其分为营养作用成分、新辅助成分、细胞毒性成分和癌症恶化促进剂。结论CME通过caspase-3/PARP和caspase-3/GSDME途径诱导A549细胞凋亡和焦亡，为CME在肿瘤患者中的临床应用提供了基础。目前的研究表明，蛹虫草提取物CME通过诱导caspase-3依赖的细胞凋亡和细胞凋亡，对A549细胞株产生凋亡和细胞凋亡。
Cordyceps militaris (CM) is traditionally used as dietary therapy for lung cancer patients in China. CM extract (CME) is hydrosoluble fraction of CM and extensively investigated. Caspase‐3‐involved cell death is considered as its major anticancer mechanism but inconclusive. Therefore, we explore its caspase‐3‐dependent programmed cell death nature (apoptosis and pyroptosis) and validate its caspase‐3‐dependent property in loss‐of‐function experiment. Component profile of CME is detected by High Performance Liquid Chromatography‐ quadrupole time‐of‐flight mass spectrometry (HPLC‐qTOF). Results show that CME causes pyroptosis‐featured cell bubbling and cell lysis and inhibits cell proliferation in A549 cell. CME induces chromatin condensing and makes PI+/annexin V+ staining in bubbling cells, indicating genotoxicity, apoptosis, and pyroptosis cell death are caused by CME. High concentration of CME (200 μg/ml) exerts G2/M and G0 cell cycles arresting and suppresses P53‐downstream proliferative proteins, including P53, P21, CDC25B, CyclinB1, Bcl‐2, and BCL2 associated agonist of cell death (BAD), but 1–100 μg/ml of CME show less effect on proteins above. Correspondingly, caspase‐3 activity and caspase‐3 downstream proteins including pyroptotic effector gasdermin‐E (GSDME) and apoptotic marker cleaved‐poly‐ADP‐ribose polymerase (PARP) are significantly promoted by CME. Moreover, regarding membrane pore formation in pyroptotic cell, expression of membrane GSDME (GSDME antibody conjugated with PE‐Cy7 for detection in flow cytometry) is remarkably increased by CME treatment. By contrast, other pyroptosis‐related proteins such as P2X7, NLRP3, GSDMD, and Caspase‐1 are not affected after CME treatment. Additionally, TET2 is unexpectedly raised by CME. In present of caspase‐3 inhibitor Ac‐DEVD‐CHO (Ac‐DC), CME‐induced cytotoxicity, cell bubbling, and genotoxicity are reduced, and CME‐induced upregulation of apoptosis (cleaved‐PARP‐1) and pyroptosis (GSDME‐NT) proteins are reversed. Lastly, 22 components are identified in HPLC‐qTOF experiment, and they are classified into trophism, neoadjuvant component, cytotoxic component, and cancer deterioration promoter according to previous references. Conclusively, CME causes caspase‐3‐dependent apoptosis and pyroptosis in A549 through caspase‐3/PARP and caspase‐3/GSDME pathways, and it provides basic insight into clinic application of CME for cancer patients. The present study has shown that the Cordyceps militaris extract CME exerts both apoptosis and pyroptosis toward A549 cell line through inducing caspase‐3‐depended pyroptosis and apoptosis.
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影响因子：
6.7
作者：
Bao, Na;Ou, Jinfeng;Chen, Li
通讯作者：
Chen, Li
影响因子：
--
作者：
Brentnall M;Rodriguez-Menocal L;De Guevara RL;Cepero E;Boise LH
通讯作者：
Boise LH
DOI：
10.1186/s40199-015-0117-6
发表时间：
2015-07-04
期刊：
Daru : journal of Faculty of Pharmacy, Tehran University of Medical Sciences
影响因子：
--
作者：
Lee HH;Lee S;Lee K;Shin YS;Kang H;Cho H
通讯作者：
Cho H
加油动物：凋亡的效应因子。
影响因子：
19
作者：
Kovacs SB;Miao EA
通讯作者：
Miao EA
影响因子：
7.9
作者：
Chou, Shang-Min;Lai, Wan-Jung;Shen, Tang-Long
通讯作者：
Shen, Tang-Long