内源性活性氧对胃癌SGC-7901细胞迁移和侵袭性的影响

Effects of endogenous reactive oxygen species on migration and invasion of gastric cancer SGC-7901 cells

目的探讨内源性活性氧(Reactive oxygen species,ROS)对胃癌SGC-7901细胞迁移和侵袭性的影响。方法采用不同浓度的脂多糖(Lipopolysaccharide,LPS)诱导细胞产生ROS,MTT法检测SGC-7901细胞存活率;荧光化学发光仪检测细胞内ROS水平;细胞划痕实验和Transwell实验检测SGC-7901细胞迁移及侵袭性的变化;Western blot检测细胞内铜锌超氧化物歧化酶(Cu/Zn-SOD)蛋白的表达。依据有无细胞内NADPH氧化酶抑制剂二苯基氯化碘盐(Diphenyleneiodonium chloride,DPI)作用于细胞,将实验分为4组:对照组、LPS组、DPI组、LPS+DPI组。所有数据采用SPSS 17.0分析。结果LPS可以促进SGC-7901细胞增殖,且LPS浓度在500ng/mL内SGC-7901细胞增殖呈现浓度和时间依耐性,LPS浓度大于500ng/mL时SGC-7901细胞增殖减弱且时间依耐性作用不明显。伴随时间推移SGC-7901细胞内ROS水平也随之增强,LPS组ROS水平、细胞划痕愈合率以及纵向迁移和侵袭穿膜的SGC-7901细胞数均高(快或多)于对照组、DPI组和LPS+DPI组,差异具有统计学意义(P均<0.01)。而LPS组Cu/Zn-SOD蛋白表达水平明显较其他组降低(P<0.01)。结论内源性ROS对胃癌SGC-7901细胞的迁移和侵袭有促进作用,通过抑制内源性ROS水平可以减缓胃癌SGC-7901细胞的迁移和侵袭。

Objective To investigate the effect of endogenous reactive oxygen species(ROS)on the migration and invasion of gastric cancer SGC-7901 cells.Methods Different concentrations of lipopolysaccharide(LPS)were used to induce cells to produce ROS,and the survival rate of SGC-7901 cells was detected by MTT assay;the intracellular ROS level was detected by fluorescence chemiluminescence;Cell scratch assay and Transwell assay were used to detect the migration and invasion of SGC-7901 cells;the protein expression of copper zinc superoxide dismutase(Cu/Zn-SOD)was detected by Western blot.Based on the presence or absence of intracellular NADPH oxidase inhibitors diphenyleneiodonium chloride(DPI)acting on the cells,the experiment was divided into four groups:control group,LPS group,DPI group and LPS+DPI group.All data were analyzed by SPSS17.0.Results LPS could promote the proliferation of SGC-7901 cells,and the proliferation of SGC-7901 cells was concentration and timedependent when the concentration of LPS was 500ng/mL,when the concentration of LPS was greater than 500ng/mL,the proliferation of SGC-7901 cells was weakened and the time-dependent effect was not obvious.Over time,the ROS levels in SGC-7901 cells also increased.The ROS level,cell scratch healing rate,and the number of SGC-7901 cells that migrated longitudinally and invaded and penetrated the membrane in the LPS group were higher(faster or more)than those in the control group,DPI group and LPS+DPI group,with statistical significances(P<0.01).The expression level of Cu/Zn-SOD protein in the LPS group was significantly lower than that in the other groups(P<0.01).Conclusion Endogenous reactive oxygen species can promote the migration and invasion of gastric cancer SGC-7901 cells.Inhibiting the level of endogenous reactive oxygen species can slow down the migration and invasion of gastric cancer SGC-7901 cells.