Effects of AZD8055 on autophagy and apoptosis in cholangiocarcinoma cells
10.3969/j.issn.1000-4718.2018.06.010
- VernacularTitle:AZD8055对胆管癌细胞自噬和凋亡的影响
- Author:
Te-Si LIU
1
;
Wen-Di YAN
;
Xue WANG
;
You LÜ
;
Ying-Shi PIAO
;
Zhen-Hua LIN
;
Xiang-Shan REN
Author Information
1. 延边大学肿瘤研究中心
- Keywords:
AZD8055;
Cholangiocarcinoma;
Autophagy;
Apoptosis
- From:
Chinese Journal of Pathophysiology
2018;34(6):1020-1024
- CountryChina
- Language:Chinese
-
Abstract:
AIM:To explore the effects of mammalian target of rapamycin (mTOR) double inhibitor AZD8055 on autophagy and apoptosis of human cholangiocarcinoma cell line HuCCT1. METHODS:The effect of AZD8055 on the viability of HuCCT1 cells was detected by MTT assay. Autophagosome was detected by acridine orange (AO) staining. Af-ter treated with AZD8055, the expression levels of apoptosis-related proteins Bcl-2, Bax and cleaved caspase-3 and auto-phagy marker proteins beclin 1, LC3 and p62 were determined by Western blot. Apoptotic rate was analyzed by flow cyto-metry with Annexin V-FITC/PI double staining. RESULTS:AZD8055 significantly inhibited the viability of HuCCT1 cells (P<0.05). AO staining showed that AZD8055 significantly increased orange granules in the cytoplasm. After treated with AZD8055, compared with the control group, the protein level of beclin 1 and the ratio of LC3-Ⅱ/LC3-Ⅰ were enhanced, while p62 was attenuated (P<0.05). The protein expression level of pro-apoptotic regulator Bax was down-regulated and anti-apoptotic regulator Bcl-2 was increased. The protein level of cleaved caspase-3 was reduced (P<0.05). The results of flow cytometry showed that AZD8055 inhibited cell apoptosis. CONCLUSION:AZD8055 inhibits the viability of cholangiocarcinoma cells, and the mechanism is closely related with autophagy induced by AZD8055.
