内皮素3/内皮素受体B对A375细胞NF-κB/ Bcl-2相关蛋白A1抗凋亡通路的调节

中华皮肤科杂志 ›› 2011, Vol. 44 ›› Issue (3): 191-194.

内皮素3/内皮素受体B对A375细胞NF-κB/ Bcl-2相关蛋白A1抗凋亡通路的调节

杨凌云¹,李艳秋²,黄巍³,黄长征⁴,陈思远⁵,曾山鹰⁶

1. 武汉华中科技大学医院皮肤科
2. 湖北省武汉市协和医院皮肤科
3. 武汉市皮肤病防治研究所
4. 华中科技大学同济医学院附属协和医院皮肤科
5. 武汉市华中科技大学同济医学院附属协和医院皮肤科
6.

收稿日期:2010-06-04 修回日期:2010-09-28 出版日期:2011-03-15 发布日期:2011-03-10
通讯作者: 黄长征 E-mail:hcz0501@126.com
基金资助:
ET3/PI3K途径在黑素瘤发病机制中的研究

Endothelin-3/endothelin receptor B（ET-3/ETRB） regulates the NF-κB/Bfl-1 anti-apoptotic pathway in a malignant melanoma cell line A375

Received:2010-06-04 Revised:2010-09-28 Online:2011-03-15 Published:2011-03-10
Contact: HUANG Chang-Zheng E-mail:hcz0501@126.com

摘要/Abstract

摘要：

目的研究内皮素3（ET-3）对人恶性黑素瘤（MM）A375细胞核因子（NF）-κB/Bfl-1抗凋亡通路的调节。方法用ET-3（100 nmol/L）刺激A375细胞24 h后，流式细胞仪检测细胞凋亡率。RT-PCR及Western印迹测定不同浓度 ET-3（0、1、10、100 nmol/L）及其与内皮素受体B（ETRB）阻断剂BQ788联合干预A375细胞后，Bfl-1在mRNA和蛋白质水平的表达。Western印迹检测相同条件干预下磷酸化NF-κB（pNF-κB）的蛋白表达。结果 ET-3可以降低A375细胞的凋亡率（F = 10.68，P < 0.05）。ET-3对A375细胞Bfl-1 mRNA和蛋白表达水平的影响呈浓度依赖性上调，BQ788明显阻断ET-3上调Bfl-1的效应（P ＜ 0.01）。Western印迹检测结果显示，ET-3显著上调pNF-κB 的表达（P ＜ 0.05），BQ788干预下其表达水平降低，接近对照组，ET-3（100 nmol/L） + BQ788组与100 nmol/L ET-3组比较，差异有统计学意义（P < 0.01）。结论 ET-3/ETRB通过激活NF-κB/Bfl-1抗凋亡通路抑制A375细胞凋亡。

关键词: NF-κB

Abstract:

Objective To investigate the modulation of ET-3 on the nuclear factor （NF）-κB/Bfl-1 anti-apoptotic pathway in a malignant melanoma cell line A375. Methods Flow cytometry was performed to detect the apoptosis in cultured A375 cells after treatment with ET-3 of 100 nmol/L for 24 hours. ET-3 of various concentrations （0, 1, 10, 100 nmol/L） was used to treat some A375 cells with or without the pretreatment with the ETRB antagonist BQ788; after another 24-hour culture, RT-PCR and Western blot were conducted to examine the mRNA expression of Bfl-1 and protein expressions of Bfl-1 and ETRB, respectively. Results The 24-hour treatment with ET-3 of 100 nmol/L significantly reduced the apoptosis rate of A375 cells （F = 10.68, P < 0.05）. The mRNA and protein expressions of Bfl-1 were up-regulated by ET-3 in a concentration dependent manner （both P < 0.01）, while BQ788 significantly blocked the ET-3-induced up-regulation（F = 420.38, 229.49, both P < 0.01）. The protein expression of pNF-κB in A375 cells was also enhanced by ET-3 of different concentrations （all P < 0.05）, but the enhancement was suppressed by BQ788, and there was a significant difference in the protein expression of pNF-κB between cells treated with ET-3 of 100 nmol/L and those treated with the combination of ET-3 of 100 nmol/L and BQ788 （F = 255.46, P < 0.01）. Conclusion ET-3/ETRB inhibits the apoptosis in A375 cells likely by activating the NF-κB/Bfl-1 anti-apoptotic pathway.

Key words: NF-kappa B

杨凌云李艳秋黄巍黄长征陈思远曾山鹰. 内皮素3/内皮素受体B对A375细胞NF-κB/ Bcl-2相关蛋白A1抗凋亡通路的调节[J]. 中华皮肤科杂志, 2011, 44(3): 191-194.

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