亚硒酸钠对胃癌SGC-7901细胞增殖、凋亡及hTERT表达的影响  

EFFECT OF SODIUM SELENITE ON THE GROWTH AND APOPTOSIS AND hTERT PROTEIN EXPRESSION OF GASTRIC CANCER SGC-7901 CELL LINE

作　　者：王慧[1] 苏衍萍[1] 葛丽[1] 刘立伟[1] 魏丽华[1] 

机构地区：[1]泰山医学院组胚教研室,泰安271000

出　　处：《营养学报》2011年第1期44-48,共5页Acta Nutrimenta Sinica

基　　金：山东省教育厅资助项目(NoJ08LG21);泰山医学院自然科学基金资助项目

摘　　要：目的探讨亚硒酸钠对人胃癌SGC-7901细胞株生长、凋亡和hTERT表达的作用及相关机制。方法用含不同浓度亚硒酸钠(0、0.5、2.5、5.0、8.0μmol/L)的培养液培养SGC-7901细胞24、48、72、96h,用相差显微镜观察亚硒酸钠对SGC-7901细胞形态的影响;MTT法检测亚硒酸钠对SGC-7901细胞增殖的影响;AnnexinV-FITC/PI双染法检测亚硒酸钠对SGC-7901细胞细胞的凋亡的影响;链霉亲和素-生物素-酶复合物(SABC)免疫细胞化学法检测亚硒酸钠对SGC-7901细胞hTERT蛋白表达的影响。结果 (1)MTT法结果显示:用亚硒酸钠培养SGC-7901细胞株24,48,72,96h后,随浓度增加,吸光度值逐渐降低,24h后当亚硒酸钠浓度大于2.5μmol/L时,与对照组比较其吸光度值均明显降低(P<0.01),48、96h后各浓度亚硒酸钠组与正常对照组比其吸光度值均明显降低(P<0.01)。随浓度的增加,亚硒酸钠对SGC-7901细胞生长抑制率逐渐增加。(2)流式细胞仪检测结果显示:亚硒酸钠可促进SGC-7901细胞发生细胞凋亡。5.0、8.0μmol/L亚硒酸钠组比正常对照组凋亡率明显升高(P<0.01)。(3)免疫细胞化学法检测结果显示:亚硒酸钠可降低SGC-7901细胞中hTERT蛋白免疫细胞化学染色的平均光密度(MOD),24h时5、8μmol/L亚硒酸钠组与对照组比MOD明显降低(P<0.01),48h时0.5、5.0、8.0μmol/L亚硒酸钠组与对照组比MOD明显降低(P<0.01),72h时2.5、5.0、8.0μmol/L亚硒酸钠组与对照组比MOD明显降低(P<0.01),96h时实验组与对照组比较MOD均明显降低(P<0.01)。结论亚硒酸钠能抑制SGC-7901细胞生长,诱导其凋亡,其作用机制可能与下调hTERT表达有关。Objective To investigate the effect and mechanism of sodium selenite on the cell growth,apoptosis and expression of human telomerase reverse transcriptase（hTERT） of human gastric cancer cell SGC-7901.Method SGC-7901 cells were treated with different concentrations of sodium selenite（0,0.5,2.5,5.0,8.0 μmol/L） at 24,48,72 and 96h,and the morphologic changes of SGC-7901 cells were observed by phase contrast microscopy.The proliferation impact of SGC-7901 cells was tested by MTT assay.SGC-7901 cells apoptosis was detected by AnnexinV-FITC,PI double staining.hTERT protein of SGC-7901 cells was stained by immunohistochemical streptavidin-biotin complex（SABC） method.Results（1）MTT assay showed that： absorbance values of SGC-7901 cells gradually decreased with the increasing concentration of sodium selenite after cultured for 24,48,72,96h.The absorbance values were significantly lower than normal control group（P0.01） when Na2SeO3 concentration was higher than 2.5μmol/L after cultured for 24h,and at all sodium selenite concentrations after culturd for 48 and 96h.The growth inhibition rate of SGC-7901 cells gradually increased with the increasing concentration of sodium selenite.（2）Flow cytometry showed that： sodium selenite facilitated SGC-7901 cells apoptosis.The apoptosis rate was significantly higher（P0.01） in 5.0,8.0 μmol/L sodium selenite group（Se group） than in the normal control group（CT group）.（3）Immunocytochemistry showed： sodium selenite reduced the mean optic density（MOD） of hTERT protein in SGC-7901 cells.The MOD was significantly lower（P0.01） in 5.0,8.0 μmol/L Se group than in CT group after cultured for 24 h in 0.5,5.0,8.0 μmol/L Se group after 48 h,in 2.5,5.0,8.0 μmol/L Se group after 72 h,and in every Se group after 96 h.Conclusion Sodium selenite can inhibit SGC-7901 cell growth and induce SGC-7901 cell apoptosis.The mechanism may be associated with reduction of hTERT expression.

关 键 词：亚硒酸钠 SGC-7901细胞株 细胞凋亡 端粒酶逆转录酶 

分 类 号：R735.2[医药卫生—肿瘤]