姜黄素对人胃癌SGC7901细胞的抑制作用及其作用机制探讨

Inhibition of curcumin on human gastric cancer SGC7901 cells and its mechanism

目的研究姜黄素对人胃癌SGC7901细胞的抑制作用及其可能作用机制。方法采用MTT比色法检测24 h和48 h时不同浓度水平(0μmol/ml、10μmol/ml、20μmol/ml、40μmol/ml、80μmol/ml)姜黄素对人胃癌SGC7901细胞生长的抑制作用;20μmol/ml姜黄素处理人胃癌SGC7901细胞24 h和48 h后,利用流式细胞术检测人胃癌SGC7901细胞的凋亡情况;Western blot法检测人胃癌SGC7901细胞中半胱氨酰天冬氨酸酶-3(caspase-3)、蛋白激酶R样内质网激酶(PERK)蛋白的表达水平;逆转录聚合酶链式反应检测人胃癌SGC7901细胞中信号转导与转录激活子1(STAT1)mRNA及信号转导与转录激活子3(STAT3)mRNA的表达量。结果对于人胃癌SGC7901细胞,姜黄素作用24 h和48 h后,细胞生长受抑制,同时药物浓度越高,作用时间越长,抑制作用越强(F分别=33.90、42.75,P均<0.05);姜黄素作用24 h、48 h后的半抑制浓度(IC50)分别为35.41μmol/ml、16.56μmol/ml;20μmol/ml姜黄素处理人胃癌SGC7901细胞24和48 h后,人胃癌SGC7901细胞的凋亡明显增加(t分别=14.58、6.27,P均<0.05)。20μmol/ml姜黄素处理人胃癌SGC7901细胞48 h后,caspase-3蛋白、PERK蛋白和STAT1 mRNA的表达明显上调(t分别=23.32、17.79、20.55,P均<0.05),而STAT3 mRNA的表达则明显下调(t=12.48,P均<0.05)。结论姜黄素通过促进人胃癌SGC7901细胞凋亡来抑制细胞增殖,具有浓度和时间依赖性;其作用机制可能与激活caspase-3凋亡通路、PERK通路和STAT信号通路等多条细胞信号通路有关。

Objective To investigate the inhibitory effect of curcumin on human gastric cancer SGC7901 cells and its mechanism.Methods MTT assay was used to detect SGC7901 cells viability after treatment with 0μmol/ml,10μmol/ml,20μmol/ml,40μmol/ml,and 80μmol/ml curcumin for 24h and 48h.After 20μmol/ml curcumin incubation for 24h and 48h in SGC7901 cells respectively,flow cytometry was employed to examine the apoptosis of SGC7901 cells.Western blotting was used to detect the expressions of caspase-3 protein and PERK protein.RT-PCR was used to detect the expressions of STAT1 mRNA and STAT3 mRNA.Results Curcumin inhibited the growth of SGC7901 cells,and the higher the drug concentration and longer time,the stronger the inhibition(F=33.90,42.75,P<0.05).The IC50 after curcumin dealing for 24h and 48h were 35.41μmol/ml and 16.56μmol/ml respectively.After 20μmol/ml curcumin incubation for 24h and 48h in SGC7901 cells respectively,the SGC7901 cells apoptosis significantly increased(t=14.58,6.27,P<0.05).After 20μmol/ml curcumin in cubation for 48h SGC7901 cells respectiuely,the expressions of caspase-3 protein,PERK protein,and STAT1 mRNA were significantly up-regulated(t=23.32,17.79,20.55,P<0.05),while the expression of STAT3 mRNA was down-regulated(t=12.48,P<0.05).Conclusion Curcumin could inhibits SGC7901 cell proliferation by promoting cell apoptosis with dose-dependence and time-dependence.The relevent mechanism of curcumin might be related to the activation of caspase-3 apoptosis pathway,PERK pathway and STAT signaling pathway.