Effect of PI3K/mTOR Signal Pathway Inhibitor XL765 on Human Leukemic KG-1 Cells.
10.19746/j.cnki.issn.1009-2137.2019.03.014
- Author:
Pin WU
1
;
Su-Ning CHEN
1
;
Qian WANG
1
;
Chuan HE
1
;
Ri ZHANG
2
Author Information
1. Department of Hematology, the First Affiliated Hospital of Soochow University, Jiangsu Institute of Hematology, Key Laboratory of Thrombosis and Hemostasis, Suzhou 215006, Jiangsu Province, China.
2. Department of Hematology, the First Affiliated Hospital of Soochow University, Jiangsu Institute of Hematology, Key Laboratory of Thrombosis and Hemostasis, Suzhou 215006, Jiangsu Province, China,E-mail: zhangri@medmail.com.cn.
- Publication Type:Journal Article
- MeSH:
Apoptosis;
Cell Line, Tumor;
Humans;
Phosphatidylinositol 3-Kinases;
Proto-Oncogene Proteins c-akt;
Quinoxalines;
Signal Transduction;
Sulfonamides;
TOR Serine-Threonine Kinases
- From:
Journal of Experimental Hematology
2019;27(3):729-734
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To explore the effect and possible mechanism of PI3K/mTOR inhibitor XL765 on KG-1 cells in vitro.
METHODS:The effect of XL765 on cell proliferation was detected by CCK-8 assay. The colony formation test (200 cells were plated in a plate for 9 days) was used to detect the effect of XL765 on the colony forming ability of KG-1 cells. The apoptosis was assessed by flow cytometry with Annexin V-FITC/PI double staining. Quantitative real-time polymerase chain reaction (q-PCR) was used to detect the expression of cell apoptosis-related genes BCL-2, BAX and caspase-3, Western blot was performed to detect the expression levels of BCL-2, BAX, Caspase-3, and the phosphorylation change of p-PI3K, p-AKT and p-S6K.
RESULTS:XL765 effectively inhibited the proliferation and the colony formation of KG-1 cells (P=0.0002). XL765 (150 nmol/L) induced KG-1 cell apoptosis (31.87±1.376%), very statistically significant different from (3.533±0.4179% ) in the control group (P＜0.01). Treatment with 150 nmol/L XL765 could in a significantly increase the expression levels of BAX and active caspase-3, and decreases expression level of the BCL-2 (P＜0.01). In accordance with these results, the Western blot further confirmed the expression decrease of BCL-2 protein along with the increase BAX and cleaved caspase-3 activity. XL765 statistically significantly down-regulated the phosphorylation levels of PI3K, AKT and S6K.
CONCLUSION:PI3K/mTOR inhibitor XL765 substantially suppresses KG-1 cell proliferation and induces apoptosis by inhibiting the activation of PI3K-AKT-mTOR signaling pathway, and regulating the apoptosis-related proteins.
