姜黄素对人肺腺癌细胞促凋亡及抗侵袭作用的研究

Apoptosis inducing and anti-invasive effects of curcumin on A549 human lung adenocarcinoma cells and its mechanism

目的 探讨姜黄素对人肺腺癌细胞 (A5 4 9)的促凋亡及抗侵袭作用。方法 采用荧光显微镜、MTT比色法、流式细胞仪(FCM)技术结合PI及AnnexinV FITC双标记染色观察姜黄素作用后 ,人肺腺癌细胞的生长及凋亡状况 ;用Westernblot法进一步验证上述结果及抗侵袭作用。结果 姜黄素作用于癌细胞后 ,荧光镜下可见细胞核破碎 ;MTT法测得不同浓度姜黄素作用后 ,可产生显著的细胞增殖抑制作用 ,IC50 =18μmol/L ;姜黄素药物浓度从 5 μmol/L增至 4 0 μmol/L ,Annexix FITC单阳性细胞 (早期凋亡细胞 )由 3 4 %增加到6 5 9% ,同时发现出现了G2 期的阻滞 ;Westernblot法观察到 10 μmol/L及 2 0 μmol/L姜黄素作用 30min后 ,出现了聚 (二磷酸腺苷 核糖 )多聚酶 (PARP)的裂解 ;姜黄素可下调MMP 2和上调TIMP 2的表达。结论 姜黄素能干扰细胞周期进程 ,对A5 4 9细胞有生长抑制作用 ,且呈剂量依赖性 ;姜黄素可能通过下调MMP 2和上调TIMP 2的蛋白表达发挥抗侵袭作用。

Objective To study the mechanism of apoptosis inducing and anti-invasive effects of curcumin on human lung adenocarcinoma cell (A549). Methods MTT colorimetry method, fluorescence mieroscopy, and FCM combining with PI and AnnexinV-FITC double pigmentation method were used to study the growth and apoptosis of A549 cells after being treated with curcumin, and Western blotting was used to identify apoptosis-inducing and anti-invasive effects. Results Under the effect of the curcumin, the nucleoli of A549 cells were found to be fragmented into different sized apoptosis bodies under fluorescence microscopy, and cell proliferation was obviously suppressed under the effect of curcumin in different concentrations, with the IC 50 value of 18μmol/L. When the curcumin concentration was increased from 5μmol/L to 40μmol/L, Annexin-FITC single positive cells (early apoptosis cell) were increased from 3.4% to 65.9%, and the proliferation of cells was blocked at G 2 phase. When curcumin concentration was increased from 10μmol/L to 20μmol/L curcumin effects 30 minutes, the expression of PARP in A549 cells was increased after 30 minutes. Curcumin could also down-regulate MMP-2 and up-regulate TIMP-2 expression. Conclusions Curcumin can interfere with cell growth cycle of A549 cells and suppress cell growth, which is concentration dependent. The anti- invasive effects of curcumin is probably the result of down-regulation of MMP-2 and up-regulation of TIMP-2 expression.