Suppression effect of expressing parvovirus H-1 nonstructural protein 1 gene on human gastric cancer cells
10.3760/cma.j.issn.0254-1432.2013.07.007
- VernacularTitle:转染及表达细小病毒H-1非结构蛋白1基因对人胃癌细胞的抑制作用
- Author:
Di ZHAO
;
Chenwen CAI
;
Jiong LIU
;
Shudong XIAO
;
Qing ZHENG
- Publication Type:Journal Article
- Keywords:
Stomach neoplasms;
H-1 parvovirus;
Viral nonstructural proteins;
Genes,tumor suppressor;
Cell aging;
Cell cycle
- From:
Chinese Journal of Digestion
2013;33(7):460-464
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the suppression effect of expressing parvovirus H-1 nonstructural protein 1 (NS1) gene on human gastric cancer cells and the possible mechanisms.Methods A recombinant enhanced green fluorescent protein (eGFP) labeled NS1 of parvovirus H-1 plasmid was constructed.Human gastric cancer cell line SGC7901 was transfected with recombinant plasmid (experiment group) or blank vector (negative control group) and blank control group was treated with equal amount of phosphate buffered saline (blank control group).After transfection,the distribution of fluorescent signal was observed under fluorescent microscope.The expression of NS1 at gene and protein level was measured.Cell growth curve of each group was drawn.The expression of cell senescence-associated β-galactosidase (SA-β-Gal) was tested.The changes of cell cycle were investigated by flowcytometry.Two groups' comparision was performed by t-test.Results After transfection,NS1 was expressed in SGC7901 cells at gene and protein level.Compared with negative control group,the fluorescent signal accumulated in cell nucleus in experiment group.The percentage of SA-β-Gal positive cell in experiment group ((30.5 ± 1.4) ％) was higher than that of negative control group ((4.4± 1.1) ％) and the difference was statistically significant (t =-12.931,P ＜ 0.01).The growth inhibition rate of SGC7901 cells from the first day to the fourth day was 45％,62％,73％ and 77％,respectively.The cell cycle of eGFP-NS1 expressed SGC7901 cells was arrested at G0/G1 phase.Conclusion Parvovirus H-1 NS1 play the role in cell nucleus of gastric cancer cell line SGC7901 and could make cell cycle arrested at G0/G1 phase,which effectively inhibited the proliferation SGC7901 cell.
