姜黄素对A549人肺腺癌细胞增殖抑制作用及其机制的研究

目的:探讨姜黄素对人肺腺癌细胞(A549)抗癌作用及其分子机制.方法:采用荧光显微镜、四甲基偶氮唑蓝(MTT)比色法、流式细胞仪(FCM)技术结合PI及AnnexinV-FITC双标记染色、Western blot法.结果:(1)姜黄素作用于癌细胞后,光镜下可见有细胞脱壁,悬浮培养液中;荧光镜下可见细胞核破碎,裂解成大小不等的凋亡小体.(2)MTT法测得不同浓度姜黄素作用A549细胞72 h后,可产生显著的细胞增殖抑制作用.将量效关系进行直线回归,得IC50值18 μmol/L.(3)姜黄素诱导凋亡作用呈剂量依赖性.随着药物浓度从5 μmo/L增加至30 μmol/L,Annexix-FITC单阳性细胞(早期凋亡细胞)由3.4%增加到59.1%;当姜黄素浓度增至40 μmol/L时,PI及AnnexinV-FITC双阳性细胞(凋亡继发性坏死细胞)成为主要的细胞组成.同时发现G2期细胞比例明显增多,出现了G2期阻滞.(4)Western blot法观察到,随着姜黄素浓度增至10 μmol/L,116 kD裂解成为89 kD片段的多聚ADP-核糖聚合酶的表达同步增加.结论:姜黄素能干扰细胞周期进程,对A549细胞有生长抑制作用,且呈剂量依赖性,其抑制作用不仅是由于非特异性的细胞毒性造成的,还与诱导肿瘤细胞发生凋亡有关.

Research of Anti-proliferation of Curcumin on A549 Human Lung Cancer Cells and Its Mechanism

OBJECTIVE: To investigate the mechanism of anti-tumor effects of curcumin on human lung cancer cell (A549). METHOD: MTT colorimetry method, fluoroscope, FCM combine PI and Annexin V-FITC double pigmentation method and Western blot method were used. RESULT: Under the effect of the curcumin, cell grew against the wall and suspended in the culture liquid, the A549 cell nucleolus were found fragmentated into different size of apoptosis body under fluoroscopy. The cell proliferation were obvious suppressed after treated with different concentration curcumin for 72 hours. The IC50 were 18 micromol/L by using linear regression. The apoptosis induced by curcumin of A549 cell is concentration dependent. With curcumin increased from 5 micromol/L to 30 micromol/L, Annexin-FITC single positive cell (early apoptosis cell) increased from 3.4% to 59.1%. When curcumin concentration reached 40 micromol/L, PI and Annexin V-FITC double positive cell (secondary apoptosis necrosis cell) became major part of cells, and the cell showed G2 phase block. Observed with western blot method, with the increase of curcumin concentration to 10 micromol/L, the expression of PARP increased simultaneously. CONCLUSION: Curcumin can interfere cell growth cycle of A549 cell and suppress cell growth. The suppression effect is concentration dependent. The effect depends not only from the nonspecific cytotoxic but also from induced cell apoptosis.