[1]地里夏提·库尔班,陈 召. Notch1调控PI3K/AKT信号通路促进黑素瘤细胞发展的研究[J].中国美容医学,2019,(01):93-96.
　Dilixiati·KUERBAN,CHEN Zhao.Notch1 Regulating PI3K/AKT Signaling Pathway Promoting the Development of Melanoma Cells[J].Medical Aesthetics and Beauty,2019,(01):93-96.
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 Notch1调控PI3K/AKT信号通路促进黑素瘤细胞发展的研究
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《中国美容医学》[ISSN:1008-6445/CN:61-1347/R]

卷:
期数:
2019年01期
页码:
93-96
栏目:
出版日期:
2019-01-15

文章信息/Info

Title:
Notch1 Regulating PI3K/AKT Signaling Pathway Promoting the Development of Melanoma Cells
文章编号:
1008-6455（2019）01-0093-04
作者:
地里夏提·库尔班; 陈 召
Author(s):
Dilixiati·KUERBAN; CHEN Zhao
关键词:
 ; [关键词]黑素瘤; Notch1; 增殖; 侵袭; 凋亡; 迁移; 信号通路
Keywords:
Key words: melanoma;  Notch1;  proliferation;  attacks;  apoptosis;  migration;  signal pathway
分类号:
R329.2
文献标志码:
A
摘要:
[摘要]目的：分析Notch1在黑素瘤发展过程中的作用和机制。方法：采用RT-PCR检测Notch1 mRNA在黑素瘤和正常黑素细胞
中的表达，采用慢病毒转染技术构建Notch1过表达的黑色素瘤细胞系，CCK法检测细胞增殖能力，Transwell检测细胞侵袭能
力，划线法检测细胞迁移能力，流式细胞术检测细胞凋亡能力，Western blot检测细胞Notch1过表达对PI3K/AKT信号通路
中AKT磷酸化水平的影响。结果：黑素瘤细胞系MM200、Mel-CV、A2058和A375细胞中Notch1 mRNA水平明显高于正常黑素细
胞系HemN-MP（P ＜0.05）。黑素瘤细胞A2058细胞中Notch1水平升高后，细胞的增殖速率显著提高，在48h、72h和96h转染
组细胞增殖能力明显高于对照组（P ＜0.05）；Transwell结果显示Notch1水平升高可以显著增加穿过小室膜A2058细胞数量
（P ＜0.05）；划线法结果显示24h转染组细胞迁移率明显高于对照组（P ＜0.05）；流式细胞检测结果显示转染组细胞早期
凋亡率明显低于对照组（P ＜0.05）。Notch1水平增加后，AKT蛋白磷酸化水平明显提高，说明Notch1能够促进PI3K/AKT信号
通路的激活。加入了Notch信号通路特异性阻断剂DAPT后，能够有效抑制Notch1过表达引起的AKT蛋白磷酸化的升高。结论：
Notch1能够调节PI3K/AKT信号通路通过促进黑素瘤的发展，本研究为临床治疗黑素瘤提供了新的思路。

Abstract:
Abstract: Objective To analyze the role and mechanism of Notch1 in the development of melanoma. Methods Notch1 mRNA
expression in normal melanocytes and melanoma was detected by RT-PCR. Slow virus transfection technique was used to
construct Notch1 expression of melanoma cell lines. Using CCK method to detect cell proliferation ability, Transwell to detect
cell invasion ability, marking method to detect cell migration ability, flow cytometry to detect cell apoptosis, Western blot
to detect cell Notch1 over expression of PI3K/AKT signal pathway of AKT phosphorylation level. Results The expression
levels of Notch1 mRNA in melanoma cell lines MM200, Mel-CV, A2058 and A375 were significantly higher than that in
normal melanocyte lines HemN-MP (P ＜0.05). When the level of Notch1 in the melanoma cell A2058 cells was increased,
the cell proliferation rate was significantly increased, and the proliferation capacity of the transfection group at 48h, 72h and
96h was significantly higher than that of the control group (P ＜0.05). Transwell results showed that increased Notch1 level
significantly increased the number of A2058 cells passing through the compartment (P ＜0.05). The results showed that the cell
migration rate in the 24h transfection group was significantly higher than that in the control group (P ＜0.05). Flow cytometry
test showed that the early apoptosis rate in transfection group was significantly lower than that in control group (P ＜0.05).
After the increase of Notch1 level, AKT protein phosphorylation level was significantly increased,suggesting that Notch1 could
promote the activation of PI3K/AKT signaling pathway.The addition of Notch signaling pathway specific blocking agent DAPT
can effectively inhibit the increase of AKT protein phosphorylation caused by Notch1 overexpression. Conclusion Notch1
can regulate PI3K/AKT signaling pathway by promoting the development of melanoma, which provides a new idea for clinical
treatment of melanoma.

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更新日期/Last Update: 2019-02-01