[1]董训虎,陈明亮,叶枫,等.氮芥通过激活腺苷酸活化蛋白激酶/自噬通路抑制角质形成细胞增殖活力[J].第三军医大学学报,2019,41(14):1295-1300.
 DONG Xunhu,CHEN Mingliang,YE Feng,et al.Nitrogen mustard inhibits keratinocyte proliferation by activating AMPK/autophagy pathway[J].J Third Mil Med Univ,2019,41(14):1295-1300.
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氮芥通过激活腺苷酸活化蛋白激酶/自噬通路抑制角质形成细胞增殖活力
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
41卷
期数:
2019年第14期
页码:
1295-1300
栏目:
基础医学
出版日期:
2019-07-30

文章信息/Info

Title:
Nitrogen mustard inhibits keratinocyte proliferation by activating AMPK/autophagy pathway
作者:
董训虎陈明亮叶枫但国蓉邹仲敏
陆军军医大学(第三军医大学)军事预防医学系防化医学教研室
Author(s):

Department of Chemical Defense, Faculty of Military Preventive Medicine, Army Medical University (Third Military Medical University),Chongqing, 400038, China

关键词:
氮芥自噬腺苷酸活化蛋白激酶角质形成细胞细胞增殖
Keywords:
nitrogen mustard autophagy AMP-activated protein kinase keratinocyte cell proliferation
分类号:
R322.991;R827.12;R827.174
文献标志码:
A
摘要:

目的 探讨腺苷酸活化蛋白激酶(AMP-activated protein kinase,AMPK)/自噬通路在氮芥(nitrogen mustard,NM)诱导角质形成细胞增殖活力下降中的作用。方法 用不同浓度(1、5、10、20 μmol/L)的氮芥单独或加入5 mmol/L自噬特异性抑制剂(3-methyladenine, 3-MA)、20 nmol/L自噬特异性激动剂(rapamycin, RAPA)、10 μmol/L AMPK特异性抑制剂(compound C, CC)处理人角质形成细胞株(HaCaT细胞)24 h后,CCK-8法检测细胞增殖活力,Western blot检测AMPK、pAMPK、LC3-B2/B1和p62的表达水平,共聚焦显微镜观察自噬体形成。结果 NM处理后HaCaT细胞增殖活力被显著抑制(P<0.05),且处理浓度越高抑制效应越明显;当NM浓度为20 μmol/L时,角质形成细胞增殖活力约下降40%。同时,NM处理能力显著提高pAMPK、LC3-B2的表达水平并下调p62的表达,且随处理浓度的增加,表达变化越明显;同时,NM可明显促进HaCaT细胞自噬体的形成。加入3-MA或CC干预后能显著抑制NM的上述效应(P<0.05),而RAPA干预对NM处理的效应无明显影响(P>0.05)。结论 氮芥能够激活AMPK/自噬通路诱导角质形成细胞的毒性损伤,抑制细胞的增殖活性。

Abstract:

Objective To explore the role of AMP-activated protein kinase (AMPK)/ autophagy signaling pathway in nitrogen mustard (NM)-induced cytotoxicity in cultured keratinocytes. Methods HaCaT cells were treated with different concentrations of NM(1, 5, 10 and 20 μmol/L) in the presence or absence of 5 mmol/L 3-methyladenine (3-MA, a specific inhibitor of autophagy), 20 nmol/L rapamycin (RAPA, a specific activator of autophagy), or 10 μmol/L compound C (a specific inhibitor of AMPK) for 24 h. CCK-8 assay was used to assess the cell viability, and the expression of p62, LC3, AMPK and pAMPK in the cells were detected using Western blotting; laser confocal microscopy was performed to detect the formation of autophagosomes in the cells after the treatments. ResultsTreatment with NM significantly decreased the viability of HaCaT cells in a dose-dependent manner, and NM at 20 μmol/L markedly decreased the cell viability to 60% of the control level. NM also dose-dependently up-regulated the phosphorylation of AMPK and LC3-B2, down-regulated the expression of p62, and significantly promoted the formation of autophagosomes in HaCaT cells. The application of 3-MA and compound C both abolished the effects of NM (P<0.05), whereas RAPA did not produce significant changes in the effect of NM (P>0.05). Conclusion NM induces cytotoxicity in cultured keratinocytes by activating AMPK/autophagy signaling pathway to inhibit the cell proliferation.

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更新日期/Last Update: 2019-07-22