姜黄素对A549人肺腺癌细胞增殖抑制作用及其机制的研究  

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

作　　者：张艰[1] 戚好文[1] 吴昌归[1] 

机构地区：[1]第四军医大学西京医院呼吸内科,陕西西安710033

出　　处：《中药材》2004年第12期923-927,共5页Journal of Chinese Medicinal Materials

基　　金：军队青年基金资助项目 (No 0 1Q12 5 );陕西省自然科学基金项目 (No 2 0 0 3C2 0 0 4)

摘　　要：目的 :探讨姜黄素对人肺腺癌细胞 (A5 4 9)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、Westernblot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A5 4 9细胞 72h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 18μmol/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 30μmol/L ,Annexix FITC单阳性细胞 (早期凋亡细胞 )由 3 4 %增加到 5 9 1% ;当姜黄素浓度增至 4 0 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 )Westernblot法观察到 ,随着姜黄素浓度增至 10 μmol/L ,116kD裂解成为 89kD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A5 4 9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 。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 IC 50 were 18 μmol/L by using linear regression The apoptosis induced by curcumin of A549 cell is concentration dependent With curcumin increased from 5μmol/L to 30μmol/L, Annexin-FITC single positive cell(early apoptosis cell) increased from 3 4% to 59 1% When curcumin concentration reached 40 μmol/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 μmol/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

关 键 词：姜黄素 FITC 人肺腺癌细胞 增殖抑制作用 剂量依赖性 A549细胞 光镜 浓度 阳性细胞 悬浮培养 

分 类 号：R734[医药卫生—肿瘤] R285[医药卫生—临床医学]