Role of Na(+)/H(+) exchanger 1 in apoptosis of HL-60 cells induced by etoposide and its mechanism.
- Author:
Yan YAN
1
;
Hua-Wen LI
;
Qing-Hua LI
;
Rong-Hua HU
;
Tian-Xiang PANG
Author Information
1. National key Laboratory of Experimental Hematology, Institute of Hematology and Hospital of Blood Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin 300020, China.
- Publication Type:Journal Article
- MeSH:
Apoptosis;
drug effects;
Cation Transport Proteins;
genetics;
metabolism;
DNA Fragmentation;
Etoposide;
pharmacology;
Gene Expression Regulation, Leukemic;
HL-60 Cells;
Humans;
RNA, Messenger;
genetics;
Sodium-Hydrogen Exchanger 1;
Sodium-Hydrogen Exchangers;
genetics;
metabolism
- From:
Journal of Experimental Hematology
2010;18(3):612-616
- CountryChina
- Language:Chinese
-
Abstract:
This study was aimed to investigate the role of Na(+)/H(+) exchanger 1 (NHE1) in apoptosis of HL-60 cells induced by etoposide. Real-time quantitative PCR (RQ-PCR) and Western blot methods were used to determine the expression of NHE1 in HL-60 cells after the treatment with etoposide. Meanwhile, laser scanning confocal microscopy was used to test the intracellular pH (pHi) of HL-60 cells. Cell apoptosis was measured by DNA fragmentation and terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assay. The results showed that etoposide induced cell apoptosis after treatment for 24 hours. The expression level of NHE1 mRNA increased by 2.848 +/- 0.886 times after treatment with etoposide for 12 hours (p < 0.01), and the expression of NHE1 protein was also up-regulated (p < 0.01). The pHi of HL-60 cells increased from 7.11 to 7.46 after treatment with etoposide for 24 hours. Treatment with cariporide could block etoposide-induced alkalinisation and enhance the apoptosis HL-60 cells. It is concluded that the expression of NHE1 is up-regulated in process of apoptosis of HL-60 cells induced by etoposide and the apoptosis depends on the pH increase caused by NHE1 higher expression.
