Mechanisms of recombinant adenovirus-mediated SD-HA fusion protein proliferation inhibition and induced apoptosis of K562 cells.
10.3760/cma.j.issn.0253-2727.2018.04.012
- Author:
Yong HUANG
;
Ping ZHANG
;
Li DU
;
Min GUI
;
Wen Li FENG
;
Zhi PENG
1
Author Information
1. Department of Infectious Diseases, Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, China.
- Publication Type:Journal Article
- Keywords:
Apoptosis;
BCR-ABL fusion protein;
Death effector domain;
Leukemia, myeloid, chronic;
Proliferation;
Src homology 2
- MeSH:
Adenoviridae;
Apoptosis;
Cell Proliferation;
Humans;
K562 Cells;
Leukemia, Myelogenous, Chronic, BCR-ABL Positive;
Oncogene Proteins, Fusion;
Phosphatidylinositol 3-Kinases
- From:
Chinese Journal of Hematology
2018;39(4):314-319
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To investigate whether fusion protein SD-HA could regulate its downstream signaling molecule activity by competing with the phospho-BCR-ABL Y177 site, and its mechanisms to inhibit proliferation and induce apoptosis of K562 cells. Methods: Co-immunoprecipitation interaction technology analysis of fusion protein SD-HA functioned by potently binding to the phospho-BCR-ABL Y177 site, Ras, MAPK and Akt activities were observed in the Ad5F35-SD-HA-treated cells. Western blot analyses of SD-HA fusion protein on cell membrane receptor pathway to death cascade caspase-8, caspase-3 and PRAP were performed. Results: Exploration into the underlying mechanisms revealed that Ad5F35-SD-HA infection functioned by binding to the phospho-BCR-ABL Y177 site, which lead to a complex with Grb2. competitively disrupted the Grb2 SH2-phospho-BCR-ABL Y177 formation. The fusion protein SD-HA could reduce the activation of Ras and phosphorylation of MAPK (p-MAPK) and the expression level of p-ELK, inhibition of Ras-MAPK signaling pathway; SD-HA fusion protein could reduce p-Akt and Akt substrate p-GSK with inhibition of PI3K-Akt signaling pathway, thereby inhibiting the proliferation of K562 cells. Caspases-8-induced apoptosis signal could be activated by DED protein binding to DED domain of precursor caspases-8. Conclusions: The strategy of fusion protein SD-HA inhibiting-Y177 BCR-ABL and Grb2 binding could be used as a novel entry point for the treatment of chronic myeloid leukemia.
