title: [Na(+)/H(+) exchanger 1 expression and its effect on apoptosis in K562 and HL-60 cells with DNA damage].
abstract: This study was aimed to investigate the expression of Na(+)/H(+) exchanger 1 (NHE1) in K562 and HL-60 cells undergoing DNA damage induced by etoposide and to elucidate the regulating mechanism. Real-time quantitative PCR (RQ-PCR) and Western blot methods were used to determine the expression of NHE1 in K562 cells after the treating with etoposide. Meanwhile, the flow cytometry was used to detect the apoptosis of leukemic cells. The luciferase reporter vector containing NHE1 promoter was constructed to measure relative luciferase activity after treating with different etoposide concentrations. The results showed that the mRNA and protein of NHE1 increased in accordance with apoptosis ratio in HL-60 cells after treated with etoposide (p < 0.05), but no such obvious increase in K562 cells. Treatment with NHE1 specific inhibitor could block etoposide induced alkalization and reduce the apoptosis ratio of HL-60 cells. The expression pattern and apoptosis alteration was not similar in K562 cells. Relative luciferase activity of reporter vector containing NHE1 promoter however increased in K562 cells after treated with etoposide. It is concluded that the expression of NHE1 is up-regulated in the process of apoptosis of HL-60 cells induced by etoposide and depends on the pHi increasing caused by NHE1 up-regulation which is not found in K562 cells although the transcriptional activity increased.
authors.affiliation: Chinese Academy of Medical Sciences, Tianjin 300020, China.
authors.forename: Hua-Wen
authors.initials: HW
authors.lastname: Li
authors.forename: Li-Hong
authors.initials: LH
authors.lastname: Wang
authors.forename: Jian
authors.initials: J
authors.lastname: Wang
authors.forename: Guo-Qiang
authors.initials: GQ
authors.lastname: Chang
authors.forename: Wei-Na
authors.initials: WN
authors.lastname: Jin
authors.forename: Ya-Ni
authors.initials: YN
authors.lastname: Lin
authors.forename: Wei
authors.initials: W
authors.lastname: Gao
authors.forename: Ruo-Jun
authors.initials: RJ
authors.lastname: Wang
authors.forename: Li
authors.initials: L
authors.lastname: Ma
authors.forename: Tian-Xiang
authors.initials: TX
authors.lastname: Pang
mesh_terms: D017209:Apoptosis; D027682:Cation Transport Proteins / Q000378:metabolism*; D004249:DNA Damage; D005047:Etoposide; D018922:HL-60 Cells; D006801:Humans; D020014:K562 Cells; D011401:Promoter Regions, Genetic; D000074082:Sodium-Hydrogen Exchanger 1; D017923:Sodium-Hydrogen Exchangers / Q000378:metabolism*
chemical_list: D027682:Cation Transport Proteins; C490998:SLC9A1 protein, human; D000074082:Sodium-Hydrogen Exchanger 1; D017923:Sodium-Hydrogen Exchangers; D005047:Etoposide