p97抑制剂NMS-873对Eca109细胞增殖、凋亡及迁移能力的影响
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摘要
目的:探讨p97抑制剂NMS-873对食管鳞状细胞癌Eca109细胞增殖、凋亡、迁移的影响。方法:用不同浓度的NMS-873分别处理Eca109细胞24、48、72 h,采用CCK-8法检测细胞增殖;在此基础上选用1、2、3μmol/L NMS-873分别作用于Eca109细胞48 h,采用Annexin V-FITC/PI双染法检测细胞凋亡,Transwell迁移实验检测细胞迁移能力的变化,Ed U荧光法检测细胞增殖。结果:NMS-873可呈时间和剂量依赖性抑制Eca109细胞增殖; 1、2、3μmol/L的NMS-873分别作用于Eca109细胞48 h后,与阴性对照组相比,随着NMS-873浓度的增加,NMS-873组Eca109细胞凋亡率提高,迁移细胞数减少,增殖率降低(P <0. 05)。结论:NMS-873能有效抑制Eca109细胞增殖,促进细胞凋亡,并降低细胞迁移能力。
Aim: To investigate the effects of p97 inhibitor NMS-873 on proliferation,apoptosis and migration of esophageal squamous cell carcinoma cell line Eca109. Methods: Eca109 cells were treated with different concentrations of NMS-873 for 24,48,and 72 hours,respectively. CCK-8 assay was used to detect proliferation of Eca109 cells. Based on this,1,2 and 3 μmol/L NMS-873 were chosen and treated Eca109 cells for 48 hours. Annexin V-FITC/PI double staining was used to detect apoptosis,Transwell migration assay was used to detect cell migration ability,and Ed U fluorescence assay was used to detect cell proliferation. Results: CCK-8 results showed that NMS-873 significantly inhibited the proliferation of Eca109 cells in a time-and dose-dependent manner( P < 0. 05). Subsequently,after NMS-873 at 1,2,and 3 μmol/L being used to treat Eca109 cells for 48 hours,compared with the negative control group,with the increase of NMS-873 concentration,the apoptosis rate of Eca109 cells in the NMS-873 groups was significantly increased,while the number of migrated cells and the proliferation rate were significantly decreased( P < 0. 05). Conclusion: NMS-873 can effectively inhibit the proliferation of Eca109 cells,promote cell apoptosis and reduce cell migration capability.
Aim: To investigate the effects of p97 inhibitor NMS-873 on proliferation,apoptosis and migration of esophageal squamous cell carcinoma cell line Eca109. Methods: Eca109 cells were treated with different concentrations of NMS-873 for 24,48,and 72 hours,respectively. CCK-8 assay was used to detect proliferation of Eca109 cells. Based on this,1,2 and 3 μmol/L NMS-873 were chosen and treated Eca109 cells for 48 hours. Annexin V-FITC/PI double staining was used to detect apoptosis,Transwell migration assay was used to detect cell migration ability,and Ed U fluorescence assay was used to detect cell proliferation. Results: CCK-8 results showed that NMS-873 significantly inhibited the proliferation of Eca109 cells in a time-and dose-dependent manner( P < 0. 05). Subsequently,after NMS-873 at 1,2,and 3 μmol/L being used to treat Eca109 cells for 48 hours,compared with the negative control group,with the increase of NMS-873 concentration,the apoptosis rate of Eca109 cells in the NMS-873 groups was significantly increased,while the number of migrated cells and the proliferation rate were significantly decreased( P < 0. 05). Conclusion: NMS-873 can effectively inhibit the proliferation of Eca109 cells,promote cell apoptosis and reduce cell migration capability.
引文
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[2] DESHAIES RJ. Proteotoxic crisis,the ubiquitin-proteasome system,and cancer therapy[J]. BMC Biol,2014,12:94
[3] DUFEY E,URRA H,HETZ C. ER proteostasis addiction in cancer biology:novel concepts[J]. Semin Cancer Biol,2015,33:40
[4] VAN DEN BOOM J,MEYER H. VCP/p97-mediated unfolding as a principle in protein homeostasis and signaling[J]. Mol Cell,2018,69(2):182
[5] XIA D,TANG WK,YE Y. Structure and function of the AAA+ATPase p97/Cdc48p[J]. Gene,2016,583(1):64
[6] ANDERSON DJ,LE MOIGNE R,DJAKOVIC S,et al. Targeting the AAA ATPase p97 as an approach to treat cancer through disruption of protein homeostasis[J]. Cancer Cell,2015,28(5):653
[7] BASTOLA P,NEUMS L,SCHOENEN FJ,et al. VCP inhibitors induce endoplasmic reticulum stress,cause cell cycle arrest,trigger caspase-mediated cell death and synergistically kill ovarian cancer cells in combination with Salubrinal[J]. Mol Oncol,2016,10(10):1559
[8] PARZYCH K,CHINN TM,CHEN Z,et al. Inadequate finetuning of protein synthesis and failure of amino acid homeostasis following inhibition of the ATPase VCP/p97[J].Cell Death Dis,2015,6:e2031
[9] MAGNAGHI P,D'ALESSIO R,VALSASINA BA,et al. Covalent and allosteric inhibitors of the ATPase VCP/p97 induce cancer cell death[J]. Nat Chem Biol,2013,9(9):U44
[10]LABBADIA J,MORIMOTO RI. The biology of proteostasis in aging and disease[J]. Annu Rev Biochem,2015,84:435
[11]CHOU TF,BULFER SL,WEIHI CC,et al. Specific inhibition of p97/VCP ATPase and kinetic analysis demonstrate interaction between D1 and D2 ATPase domains[J]. J MolBiol,2014,426(15):2886
[12]XUE L,BLYTHE EE,FREIBERGER EC,et al. Valosincontaining protein(VCP)-adaptor interactions are exceptionally dynamic and subject to differential modulation by a VCP inhibitor[J]. Mol Cell Proteomics,2016,15(9):2970
[13]NOI K,YAMAMOTO D,NISHIKORI S,et al. Highspeed atomic force microscopic observation of ATP-dependent rotation of the AAA+chaperone p97[J]. Struc-ture,2013,21(11):1992
[14]FRANZ A,ACKERMANN L,HOPPE T. Create and preserve:proteostasis in development and aging is governed by Cdc48/p97/VCP[J]. Biochim Biophys Acta,2014,1843(1):205
[15]SKROTT Z,MISTRIK M,ANDERSEN KK,et al. Alcoholabuse drug disulfiram targets cancer via p97 segregase adaptor NPL4[J]. Nature,2017,552(7684):194