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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 growth, apoptosis and hTERT expression of human gastric cancer cell line SGC-7901. Methods SGC-7901 cells were cultured in culture medium containing different concentrations of sodium selenite (0,0.5,2.5,5.0,8.0μmol / L) for 24,48,72,96h. The effects of sodium selenite on SGC- 7901 cells; the effect of sodium selenite on the proliferation of SGC-7901 cells was detected by MTT assay; the apoptosis of SGC-7901 cells was detected by Annexin V-FITC / PI double staining; And SABC immunocytochemistry to detect the effect of sodium selenite on hTERT protein expression in SGC-7901 cells. Results (1) MTT assay showed that after 24, 48, 72 and 96 hours of culture with sodium selenite, SGC-7901 cells decreased gradually with increasing concentration of sodium selenate. When sodium selenite concentration was more than 2.5μmol / (P <0.01). Compared with the control group, the absorbance values of sodium selenite group and the normal control group were significantly decreased after 48 and 96 hours (P <0.01). With the increase of concentration, the inhibitory rate of sodium selenite on SGC-7901 cells increased gradually. (2) The results of flow cytometry showed that sodium selenite can promote the apoptosis of SGC-7901 cells. The apoptosis rates in 5.0 and 8.0μmol / L sodium selenite group were significantly higher than those in normal control group (P <0.01). (3) Immunocytochemistry results showed that sodium selenite could decrease the average optical density (MOD) of hTERT protein immunocytochemical staining in SGC-7901 cells, and at 5 and 8 μmol / L sodium selenite group, (P <0.01). Compared with MOD, the SOD of 0.5, 5.0, 8.0μmol / L sodium selenite group decreased significantly at 48h (P <0.01) The MOD of sodium selenate group and control group were significantly lower than that of the control group (P <0.01). Compared with the control group, the MOD of the sodium selenate group was significantly decreased at 96 hours (P <0.01). Conclusion Sodium selenite can inhibit SGC-7901 cell growth and induce its apoptosis, and its mechanism may be related to the down-regulation of hTERT expression.