Inhibition of deubiquitination by PR-619 induces apoptosis and autophagy via ubi-protein aggregation-activated ER stress in oesophageal squamous cell carcinoma.
Inhibition of deubiquitination by PR-619 induces apoptosis and autophagy via ubi-protein aggregation-activated ER stress in oesophageal squamous cell carcinoma.
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PR-619 抑制去泛素化通过泛蛋白聚集激活的内质网应激诱导食管鳞状细胞癌凋亡和自噬
DOI：
10.1111/cpr.12919
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发表时间：
2021-01
影响因子：
8.5
通讯作者：
Chen P
中科院分区：
文献类型：
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作者：
Wang L;Li M;Sha B;Hu X;Sun Y;Zhu M;Xu Y;Li P;Wang Y;Guo Y;Li J;Shi J;Li P;Hu T;Chen P
靶向去泛素化酶（DUBs）已成为抗癌药物开发的一个有前途的途径。然而，pan‐DUB抑制剂PR‐619对食管鳞状细胞癌（ESCC）细胞的作用和机制仍有待研究。通过CCK-8和PI染色评价PR-619对ESCC细胞生长和细胞周期的影响。Annexin V-FITC/PI双染色检测细胞凋亡。LC 3免疫荧光染色和吖啶橙子染色检测自噬。通过Fluo-3AM荧光监测细胞间Ca 2+浓度。通过免疫印迹法测定ubi蛋白的蓄积和内质网（ER）应激相关蛋白和CaMKKβ-AMPK信号传导的表达。PR-619可通过下调cyclin B1和上调p21的表达，抑制食管鳞癌细胞生长，并诱导其发生G2/M期阻滞。同时，PR-619导致泛素化蛋白的积累，诱导ER应激并通过ATF 4-Noxa轴触发细胞凋亡。此外，内质网应激还可通过Ca 2 +-CaMKKβ-AMPK信号通路增加胞浆Ca 2+，进而刺激自噬。Ubiquitin E1抑制剂PYR-41可以减少Ubi-proteins的积累，减轻ER应激、G2/M细胞周期阻滞、凋亡和自噬。此外，通过氯喹或巴弗洛霉素A1阻断自噬增强了PR-619诱导的细胞生长抑制作用和凋亡。我们的研究结果揭示了一般DUBs抑制剂（PR-619）的细胞毒性作用的未被认识的机制，并暗示靶向DUBs可能是一种潜在的抗ESCC策略。PR-619可通过下调cyclin B1的表达和上调p21蛋白水平抑制食管鳞癌细胞生长，并诱导细胞周期阻滞于G2/M期。同时，PR-619处理导致泛素化蛋白的积累，诱导ER应激并通过ATF 4-Noxa轴触发凋亡。此外，内质网应激还可通过Ca 2 +-CaMKKβ-AMPK信号通路增加胞浆内Ca 2+，进而刺激细胞自噬。
Targeting the deubiquitinases (DUBs) has become a promising avenue for anti‐cancer drug development. However, the effect and mechanism of pan‐DUB inhibitor, PR‐619, on oesophageal squamous cell carcinoma (ESCC) cells remain to be investigated. The effect of PR‐619 on ESCC cell growth and cell cycle was evaluated by CCK‐8 and PI staining. Annexin V‐FITC/PI double staining was performed to detect apoptosis. LC3 immunofluorescence and acridine orange staining were applied to examine autophagy. Intercellular Ca2+ concentration was monitored by Fluo‐3AM fluorescence. The accumulation of ubi‐proteins and the expression of the endoplasmic reticulum (ER) stress‐related protein and CaMKKβ‐AMPK signalling were determined by immunoblotting. PR‐619 could inhibit ESCC cell growth and induce G2/M cell cycle arrest by downregulating cyclin B1 and upregulating p21. Meanwhile, PR‐619 led to the accumulation of ubiquitylated proteins, induced ER stress and triggered apoptosis by the ATF4‐Noxa axis. Moreover, the ER stress increased cytoplasmic Ca2+ and then stimulated autophagy through Ca2+‐CaMKKβ‐AMPK signalling pathway. Ubiquitin E1 inhibitor, PYR‐41, could reduce the accumulation of ubi‐proteins and alleviate ER stress, G2/M cell cycle arrest, apoptosis and autophagy in PR‐619‐treated ESCC cells. Furthermore, blocking autophagy by chloroquine or bafilomycin A1 enhanced the cell growth inhibition effect and apoptosis induced by PR‐619. Our findings reveal an unrecognized mechanism for the cytotoxic effects of general DUBs inhibitor (PR‐619) and imply that targeting DUBs may be a potential anti‐ESCC strategy. PR‐619 could inhibit oesophageal squamous cell carcinoma cell growth and induce G2/M cell cycle arrest by downregulating the expression of cyclin B1 and upregulating the protein level of p21. Meanwhile, PR‐619 treatment led to the accumulation of ubiquitylated proteins, induced ER stress and triggered apoptosis by ATF4‐Noxa axis. Moreover, the ER stress increased cytoplasmic Ca2+ and then stimulated autophagy through Ca2+‐CaMKKβ‐AMPK signaling pathway.
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自噬和neddylation途径的协同抑制作为靶向肝癌的新型治疗方法
影响因子：
--
作者：
Chen P;Hu T;Liang Y;Jiang Y;Pan Y;Li C;Zhang P;Wei D;Li P;Jeong LS;Chu Y;Qi H;Yang M;Hoffman RM;Dong Z;Jia L
通讯作者：
Jia L
Neddylation 抑制通过 ATF4-CHOP-DR5 轴激活人食管癌细胞的外源性凋亡途径
影响因子：
11.5
作者：
Chen, Ping;Hu, Tao;Jia, Lijun
通讯作者：
Jia, Lijun
影响因子：
6
作者：
Jacomin AC;Taillebourg E;Fauvarque MO
通讯作者：
Fauvarque MO
影响因子：
6
作者：
Kuo, Kuan-Lin;Liu, Shing-Hwa;Huang, Kuo-How
通讯作者：
Huang, Kuo-How
DOI：
10.1158/1078-0432.ccr-13-2658
发表时间：
2014-06-15
期刊：
Clinical cancer research : an official journal of the American Association for Cancer Research
影响因子：
--
作者：
Coughlin K;Anchoori R;Iizuka Y;Meints J;MacNeill L;Vogel RI;Orlowski RZ;Lee MK;Roden RB;Bazzaro M
通讯作者：
Bazzaro M