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目的探讨不同浓度姜黄素诱导人肺腺癌A549细胞凋亡中Orai1和STIM1mRNA水平的改变。方法 A549细胞分别暴露于5、10、15、20、25和30μmol/L姜黄素12、24和48 h后,应用四甲基偶氮唑蓝(MTT)比色法测定细胞增殖活力;将不同浓度姜黄素孵育A549细胞24 h后,流式细胞仪检测细胞凋亡率;real-time PCR确定25μmol/L姜黄素处理24 h后,Orai1和STIM1mRNA表达水平。结果姜黄素对A549细胞的抑制作用呈剂量依赖性和时间依赖性,不同浓度姜黄素处理A549细胞24 h细胞存活率分别为96.5%、95.0%、77.4%、63.9%、57%、39.6%,与对照组相比,差异有统计学意义(P<0.05或P<0.01)。姜黄素处理后,流式细胞仪分析结果显示,细胞凋亡率呈明显剂量依赖关系(P<0.05或P<0.01)。经姜黄素处理后A549细胞Orai1和STIM1mRNA表达量均明显低于对照组(P<0.01)。结论姜黄素可能引起A549细胞Orai1和STIM1 mRNA表达水平发生改变,通过下调Orai1和STIM1mRNA表达量引起细胞凋亡。
Objective To investigate the changes of Orai1 and STIM1 mRNA in human lung adenocarcinoma A549 cells induced by different concentrations of curcumin. Methods A549 cells were exposed to 5, 10, 15, 20, 25 and 30 μmol / L curcumin for 12, 24 and 48 h, respectively. Cell viability was measured by MTT assay. The apoptosis rate of A549 cells was detected by flow cytometry (FCM) after the cells were incubated with curcumin for 24 hours. The mRNA expression levels of Orai1 and STIM1 were determined by real-time PCR after treated with 25μmol / L curcumin for 24 hours. Results Curcumin had a dose-dependent and time-dependent inhibitory effect on A549 cells. The survival rates of cells treated with different concentrations of curcumin for 24 h were 96.5%, 95.0%, 77.4%, 63.9%, 57%, 39.6% Compared with the control group, the difference was statistically significant (P <0.05 or P <0.01). After curcumin treatment, the results of flow cytometry showed that apoptosis rate showed a significant dose-dependent (P <0.05 or P <0.01). The mRNA expressions of Orai1 and STIM1 in A549 cells after curcumin treatment were significantly lower than those in control group (P <0.01). Conclusion Curcumin may induce the alteration of Orai1 and STIM1 mRNA expression in A549 cells and induce apoptosis by down-regulating the expression of Orai1 and STIM1 mRNA.