[1]龙剑文,罗晶,尹绪文.重楼皂苷Ⅰ通过抑制PI3K/Akt/mTOR通路诱导人黑素瘤A375细胞凋亡与自噬[J].中国皮肤性病学杂志,2019,(08):863-868.[doi:10.13735/j.cjdv.1001-7089.201812158]
 LONG Jianwen,LUO Jing,YIN Xuwen.Induces the Autophagy and Apoptosis of Melanoma A375 Cells by Suppressing PI3K/Akt/mTOR Signaling Pathway[J].The Chinese Journal of Dermatovenereology,2019,(08):863-868.[doi:10.13735/j.cjdv.1001-7089.201812158]
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重楼皂苷Ⅰ通过抑制PI3K/Akt/mTOR通路诱导人黑素瘤A375细胞凋亡与自噬
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《中国皮肤性病学杂志》[ISSN:1001-7089/CN:61-1197/R]

卷:
期数:
2019年08期
页码:
863-868
栏目:
论著
出版日期:
2019-07-15

文章信息/Info

Title:
Induces the Autophagy and Apoptosis of Melanoma A375 Cells by Suppressing PI3K/Akt/mTOR Signaling Pathway
文章编号:
1001-7089(2019)08-0863-06
作者:
龙剑文12罗晶2尹绪文2
1.湖北中医药大学第一临床学院,湖北 武汉 430061; 2.湖北省中医院皮肤科,湖北 武汉 430061
Author(s):
LONG Jianwen12LUO Jing2YIN Xuwen2
(1.The First Clinical Medical School of Hubei University of Chinese Medicine,Wuhan 430061,China; 2.Department of Dermatology,Hubei Provincial Hospital of Chinese Medicine,Wuhan 430061,China)
关键词:
重楼皂苷Ⅰ PI3K/Akt/mTOR通路 黑素瘤 凋亡 自噬
Keywords:
Polyphylin Ⅰ PI3K/Akt/mTOR signal pathway Melanoma Apoptosis Autophagy
分类号:
R 739.5
DOI:
10.13735/j.cjdv.1001-7089.201812158
文献标志码:
A
摘要:
目的 探讨重楼皂苷Ⅰ(Polyphylin Ⅰ,PPⅠ)对人黑素瘤A375细胞凋亡和自噬的影响以及相关机制,确定PPⅠ诱导的细胞自噬与其促进A375细胞凋亡的相关性。方法 PPⅠ处理 A375 细胞后,采用CCK-8法检测细胞活力; Annexin V-FITC /PI 双标法检测PPⅠ 对人黑素瘤细胞凋亡的作用; 免疫印迹法检测相关蛋白的表达水平; 通过观察细胞内自噬标志分子LC3的分布与表达来确定PPⅠ 对自噬的诱导作用; 采用氯喹抑制细胞自噬后,观察PPⅠ对细胞凋亡的影响。结果 PPⅠ能抑制A375细胞增殖活力; PPⅠ抑制PI3K/Akt/mTOR通路的活化并促进了A375细胞内 Bax 和 cleaved-caspase 3的表达,抑制Bcl-2表达,诱导人黑素瘤细胞发生凋亡; PPⅠ促进了A375细胞LC-Ⅱ/LC-Ⅰ值和Beclin-1蛋白表达的升高,下调了p62蛋白的水平,促进了细胞自噬; PI3K/Akt/mTOR通路活化剂IGF-1处理A375细胞后抑制了PPⅠ的上述作用; PPⅠ与自噬抑制剂氯喹联用后,A375细胞凋亡数量明显减弱。结论 PPⅠ 能抑制A375细胞的增殖,诱导细胞发生凋亡与自噬,抑制自噬减少了PPⅠ 诱导的人黑素瘤细胞凋亡。
Abstract:
Objective To investigate the effect of Polyphylin Ⅰ on the apoptosis and autophagy in human melanoma A375 cells line via PI3K/Akt/mTOR signal pathway.Methods The cell growth was determined by CCK-8 assay,and cell apoptosis was analyzed by Annexin V-FITC apoptosis detection kit.The protein expressions were examined by Western blot analysis,and the positive expression of LC3 in A375 cells were detected by immunofluorescence staining.Results Polyphylin Ⅰ inhibited the A375 cells viability in a dose-dependent manner.Polyphylin Ⅰ suppressed the activation of PI3K/Akt/mTOR signal pathway in A375 cells,and also induced apoptosis of A375 cells through reducing the expression of Bcl-2 and enhancing the expression of Bax and cleaved-caspase-3.The enhanced green fluorescence intensity of LC3 was observed in A375 cells in a dose-dependent manner.Polyphylin Ⅰ treatment promoted A375 cells autophagy,increased the expression of Beclin-1 and the value of LC3-Ⅱ/ LC3-Ⅰ,and reduced the expression of p62.However,treatment with insulin-like growth factor 1(IGF-1)reversed these Polyphylin Ⅰ-mediated effects on the activation of PI3K/Akt/mTOR signal pathway,cell apoptosis and autophagy; meanwhile,suppression of autophagy by chloroquin,an inhibitor of autophagy,decreased the number of apoptotic cells.Conclusion Polyphylin Ⅰ inhibits cell proliferation and triggers apoptosis and autophagy in human melanoma A375 cells via inhibiting PI3K/Akt/mTOR signal pathway.In addition,the blocking autophagy can alleviate the Polyphylin Ⅰ-induced melanoma cell apoptosis.

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[2]龙剑文,皮先明,涂亚庭.重楼皂苷Ⅰ对IL-17刺激HaCaT细胞分泌VEGF和IL-23的影响[J].中国皮肤性病学杂志,2016,(05):453.[doi:10.13735/j.cjdv.1001-7089.201509086]
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备注/Memo

备注/Memo:
[基金项目] 武汉市中青年医学骨干人才培养项目; 中央级公益性科研院所基本科研业务费专项资金资助(YZ1839)
[作者单位] 1.湖北中医药大学第一临床学院,湖北 武汉 430061; 2.湖北省中医院皮肤科,湖北 武汉 430061
更新日期/Last Update: 2019-07-15