摘要
PI3K/AKT信号通路的异常与肿瘤的发生、转移具有非常密切的关系,在乳腺癌细胞中更为常见。该研究从自主设计合成的小分子样品库中筛选出可有效诱导乳腺癌细胞BT549凋亡的新甲磺酰胺类肿瘤抑制剂DHW-51。选择PTEN/p53双基因敲除的人乳腺癌细胞株BT549,通过细胞增殖实验发现,DHW-51可抑制BT549细胞增殖并且呈剂量和时间依赖关系。溶血实验结果表明,DHW-51浓度为40μmol/L时,血红细胞的溶血活性低于10%,DHW-51对正常细胞没有毒性作用。此外,流式细胞术实验证明了DHW-51可以诱导BT549细胞凋亡,DHW-51浓度为20μmol/L时caspase3的活性增加,pro-caspase3蛋白表达下降。Western blot结果显示,BT549细胞中AKT的磷酸化水平显著下降,但AKT总蛋白的表达水平未受影响,证明DHW-51诱导BT549细胞凋亡的靶点可能是AKT蛋白,提示DHW-51可能是以AKT为靶点的肿瘤抑制剂。
The abnormal PI3K/AKT signaling pathway accounts for tumorigenesis, metastasis and multidrug resistance, especially in breast cancer. In this article, we discovered a new sulfonamide tumor inhibitor, DHW-51, which induced the apoptosis in breast cancer cell line BT549. When p53 and PTEN were both knockdown in BT549, it showed that DHW-51 effectively inhibited the survival of BT549 cell in both dose-and time-dependent manner. The hemolytic test result indicated that DHW-51 had inconsiderable effect on normal cells when hemolytic activity of red blood cell showed less than 10% with 40 μmol/L of DHW-51. In addition, FACS results also suggested that DHW-51 induced apoptosis of BT549 cells significantly. They used flow cytometry to show that DHW-51 could significantly induce apoptosis of BT549 cells, the activity of caspase3 increased with 20 μmol/L of DHW-51, however, the expression of pro-caspase3 protein decreased. As showed in the results of Western blot, the phosphorylation level of AKT in BT549 cells decreased significantly, nevertheless, the expression level of the AKT protein was not affected. We speculated that DHW-51 induced apoptosis of BT540 by targeting AKT, and suggested DHW-51 was an AKT-targeted tumor inhibitor.
作者
王冠
柴少蒙
Seria-Masole Shonyela
王明玥
郝明月
宋宏锐
施维
Wang Guan;Chai Shaomeng;Seria.Masole Shonyela;Wang Mingyue;Hao Mingyue;Song Hongrui;Shi Wei(IFaeu!ty of Agronomy,Jilin Agricultural University,Changehun 130118,China;College of Life Sciences,Jilin University,Changchun 130118,China;College of Animal Science and Technology,Jilin Agricultural University,Changchun 130118,China;Ministry of Education,Shenyang Pharmaceutical University,Shenyang 1100i 6,China)
出处
《中国细胞生物学学报》
CAS
CSCD
2018年第9期1537-1546,共10页
Chinese Journal of Cell Biology
基金
吉林农业大学校内启动基金资助的课题~~
关键词
甲磺酰胺
AKT
凋亡
人乳腺癌
sulfonamide
AKT
apoptosis
human breast cancer
作者简介
通讯作者:王冠,Tel:0431-84533277,E-mail:wxkzbwg@163.com
