Zhu Qiaohua, Luo Meihua, Zhou Chengyu, Chen Zhixian, Huang Wei, Huang Jiangyuan, Zhao Shufeng, Yu Xinfa
Department of Oncology, Shunde Hospital of Southern Medical University, Shunde 528308, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2018 Dec 30;38(12):1476-1484. doi: 10.12122/j.issn.1673-4254.2018.12.13.
To investigate the effect of danusertib (Danu), an inhibitor of Aurora kinase, on the proliferation, cell cycle, apoptosis, and autophagy of hepatocellular carcinoma HepG2 cells and explore the underlying mechanisms.
MTT assay was used to examine the effect of Danu on the viability of HepG2 cells to determine the IC50 of Danu. The effect of Danu on cell cycle distribution, apoptosis and autophagy were determined using flow cytometry. Western blotting was used to detect the expressions of the proteins related to cell cycle, apoptosis and autophagy. Chloroquine was used to suppress Danuinduced autophagy to test the apoptosis-inducing effect of Danu.
Danu significantly inhibited the proliferation of HepG2 cells with IC of 39.4 μmol and 14.4 μmol at 24 h and 48 h, respectively. Danu caused cell cycle arrest in G/M phase in HepG2 cells and led to polyploidy accumulation via up-regulating the expressions of p53 and p21 and down-regulating the expressions of cyclin B1 and DC2. Danu also caused apoptosis of HepG2 cells through up-regulating the expressions of Bax, Puma, cleaved caspase-3, cleaved caspase-9, cleaved PARP and cytochrome C and down-regulating the expressions of Bcl-xl and Bcl-2. Danu induced autophagy via activating AMPK signaling and inhibiting PI3K/PTEN/AKT/mTOR axis, and inhibition of Danu-induced autophagy with chloroquine enhanced the pro-apoptotic effect of Danu.
Danu inhibits cell proliferation and induces cell cycle arrest in G/M phase, apoptosis and cytoprotective autophagy in HepG2 cells.
研究极光激酶抑制剂达努昔布（Danu）对肝癌HepG2细胞增殖、细胞周期、凋亡及自噬的影响，并探讨其潜在机制。
采用MTT法检测Danu对HepG2细胞活力的影响，以确定Danu的半数抑制浓度（IC50）。采用流式细胞术检测Danu对细胞周期分布、凋亡和自噬的影响。采用蛋白质印迹法检测与细胞周期、凋亡和自噬相关蛋白的表达。使用氯喹抑制Danu诱导的自噬，以检测Danu的促凋亡作用。
Danu显著抑制HepG2细胞的增殖，在24小时和48小时时的IC50分别为39.4 μmol和14.4 μmol。Danu导致HepG2细胞的细胞周期阻滞于G/M期，并通过上调p53和p21的表达以及下调细胞周期蛋白B1和DC2的表达导致多倍体积累。Danu还通过上调Bax、Puma、裂解的半胱天冬酶-3、裂解的半胱天冬酶-9、裂解的聚（ADP-核糖）聚合酶和细胞色素C的表达以及下调Bcl-xl和Bcl-2的表达，导致HepG2细胞凋亡。Danu通过激活AMPK信号并抑制PI3K/PTEN/AKT/mTOR轴诱导自噬，用氯喹抑制Danu诱导的自噬增强了Danu的促凋亡作用。
Danu抑制HepG2细胞的增殖，并诱导其细胞周期阻滞于G/M期、凋亡和细胞保护性自噬。
