黄芩素通过抑制PI3K/AKT/mTOR通路诱导膀胱癌细胞凋亡

doi:10.6040/j.issn.1671-7554.0.2018.1267

摘要/Abstract

摘要： 目的研究黄芩素对膀胱癌细胞的作用及机制。方法膀胱癌细胞T24和5637经黄芩素(处理组)或二甲基亚砜(对照组)处理后,3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)以及胸腺嘧啶核苷类似物(EdU)插入实验检测细胞增殖和DNA复制速率;划痕实验和Transwell实验检测细胞的迁移和侵袭能力;流式细胞技术检测细胞周期和细胞凋亡水平;Western blotting检测PI3K/AKT/mTOR信号通路蛋白、B淋巴细胞瘤-2(Bcl-2)和B淋巴细胞瘤-2相关X蛋白(Bax)等蛋白的表达水平;裸鼠成瘤实验检测黄芩素对于体内膀胱癌细胞增殖能力的影响。结果黄芩素可以在体内和体外抑制膀胱癌细胞的增殖速率,高浓度黄芩素体外抑制率可达80%以上,体内抑制率约50%。黄芩素可以减缓DNA复制并降低肿瘤细胞的迁移和侵袭能力,诱导细胞停滞在G₀/G₁期,发生细胞凋亡,与对照组相比差异有统计学意义。黄芩素处理后,PI3K的表达以及AKT和mTOR的磷酸化水平降低,Bax/Bcl-2的比值升高。结论黄芩素通过抑制PI3K/AKT/mTOR信号通路抑制膀胱癌细胞增殖,发生周期阻滞,诱导细胞凋亡。

关键词: 黄芩素, 膀胱癌, B淋巴细胞瘤-2, PI3K/AKT/mTOR, 凋亡

Abstract: Objective To explore the effects of baicalein on bladder cancer cells and the underlying mechanism. Methods Bladder cancer cells, T24 and 5637, were treated with baicalein(the treatment group)or dimethyl sulfoxide(the control group). The rate of cell proliferation and DNA replication were examined by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide(MTT)assays and 5-Ethynyl-2'-deoxyuridine(EdU)incorporation assays. The migration and invasion of cancer cells were explored by Wound healing assays and transwell assay. The cell cycle and cell apoptosis were detected by flow cytometry. The protein levels of PI3K/AKT/mTOR, Bcl-2, Bax and so on were explored by Western blotting. The effects of baicalein on cell proliferation in vivo were explored by tumor formation on nude mice. Results The inhibition rate of baicalein on cell proliferation was above 80% in vitro and about 50% in vivo. Baicalein could interfere DNA replication and suppress the migration and invasion of bladder cancer cells. Baicalein could also induce G₀/G₁ phase arrest and apoptosis of bladder cancer cells. Treatment of baicalein down-regulated the expression of PI3K and inhibit the phosphorylation of AKT and mTOR, with an increase in Bax/Bcl-2 ratio. Conclusion Baicalein can inhibit cell proliferation, induce cell cycle arrest and apoptosis of bladder cancer cells via suppression of the PI3K/AKT/mTOR pathway.

Key words: Baicalein, Bladder cancer, B-cell lymphoma 2, PI3K/AKT/mTOR, Apoptosis

中图分类号:

R737.14

王健,李健,王勇,朱耀丰. 黄芩素通过抑制PI3K/AKT/mTOR通路诱导膀胱癌细胞凋亡[J]. 山东大学学报 (医学版), 2019, 57(9): 74-82.

WANG Jian, LI Jian, WANG Yong, ZHU Yaofeng. Baicalein induces apoptosis of bladder cancer cells via inhibition of the PI3K/AKT/mTOR pathway[J]. Journal of Shandong University (Health Sciences), 2019, 57(9): 74-82.

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