Effects of histidine triad nucleotide-binding protein 1(HINT1)on the proliferation and apoptosis of human melanoma cell line A375
10.3760/cma.j.jssn.0412-4030.2010.01.006
- VernacularTitle:HINT1对人黑素瘤细胞A375增殖和凋亡的影响
- Author:
Yang GUAN
;
Yiqun JIANG
;
Ying GAO
;
Hao CHEN
;
Xuesi ZENG
;
Jianfang SUN
- Publication Type:Journal Article
- Keywords:
Tumor cells,cultured;
Melanoma;
Cell proliferation;
Apoptosis;
HINT1
- From:
Chinese Journal of Dermatology
2010;43(1):10-13
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the effects of HINT1 on the proliferation and apoptosis of human melanoma cell line A375.Methods Three cell clones were used in the experiment.including A375 cells that were previously transfected with eukaryotic expressing vector pcDNA.3.1/myc-His(-)A-HINT1 and highly expressed HINTI(HINT1-A375),A375 cells transfected with pcDNA.3.1/myc-His(-)A empty vector(neo-A375)and untransfected A375 cells.M3T assay,flow cytometry and terminal deoxynucleotjdyl transferase (TdT)-mediated dUTP nick end labeling(TUNEL)were performed to detect the proliferation,cell cycle and apoptosis of the three kinds of cells,respectively.The relative activity of Caspase 3,8 and 9 was measured by spectrophotometry,and the protein expression of Bcl-2,Bax,Cytoehrome C and p53 by Western blot. Results Compared with neo-A375 and untransfected A375 cells,HINT1-A375 cells grew more slowly(P<0.05)with an increase in G1-phase population(73.17%±3.99%,F=25.65,P<0.05).a decrease in S-phase population(16.75%±1.62%,F=75.48,P<0.01),and a pronounced late apoptosis(23.57%±9.58%,F=11.71,P<0.01).A significant increase was also observed in the percentage of apoptotic cells(12%±1%,F=358.02,P<0.01)as well as in the relative activity of Caspase 9(0.45±0.03,F=135.62,P<0.01)and Caspase 3(0.46±0.04,F=90.28,P<0.01)in HINT1-A375 cells compared with the other two kinds of cells.Western blot showed upregulated expressions of Bax,Cytochrome C and p53 but downregulated expres-sion of Bel-2 in HINT1-A375 cells.Conclusions The overexpression of HINT1 could inhibit the proliferation and promote the apoptosis of A375 cells with cell cycle arrest in G1 phase,hinting that HINT1 may be a tumor suppressor gene in human melanoma.
