姜黄素抗人肺腺癌A549/DDP细胞增殖及诱导凋亡的作用

目的 研究姜黄素对人肺腺癌A549/DDP细胞增殖的抑制作用及其对细胞凋亡的影响.方法 采用四甲基偶氮唑蓝(MTT)测定细胞对顺铂的敏感性和姜黄素对细胞增殖的抑制作用;倒置相差显微镜和荧光显微镜下观察细胞形态学的变化;应用原位末端标记(TUNEL)染色及流式细胞仪检测细胞凋亡情况;以Western blot分析凋亡相关蛋白表达变化.结果 实验中所用的A549/DDP细胞对顺铂耐药,耐药指数为10.82.姜黄素对A549及A549/DDP细胞增殖均有抑制作用,并且呈时间、浓度依赖性(P＜0.05),对两细胞株的半数抑制浓度分别为16.28/μmol/L和18.06μmol/L,差异无统计学意义(P＞0.05).姜黄素处理A549/DDP细胞后,细胞变形、缩小、脱落.Hoechst染色显示A549/DDP细胞核缩小、变形,致密浓染,荧光成团块分布.TUNEL法以及流式细胞仪分析结果示细胞凋亡率呈明显剂量依赖关系(P＜0.05).Western blot结果显示,随姜黄素浓度的增加,天冬氨酸特异性半胱氨酸蛋白酶8(cysteinyl aspartate-specific protease-8,caspase-8)p10蛋白水平增加.结论 A549/DDP细胞对姜黄素无抗药性,姜黄素对人肺腺癌A549/DDP细胞增殖具有抑制作用,并能诱导细胞凋亡.caspase-8的活化是其中的机制之一.

Anti-proliferation and inducing apoptosis effects of curcumin on human lung adenocarcinoma cell line A549/DDP

Objective To investigate the inhibitory effect of curcumin on the human lung adenocarcinoma cell line A549/DDP and its effect on apoptosis.Methods The inhibitory effects of cisplatin and curcumin on cell proliferation were tested by MTT assay.The morphologic changes of tumor tissue were observed under inverted phase contrast microscope and fluorescence microscope.The apoptotic state of A549/DDP was measured by TUNEL and flow cytometry.Western-blot was used to determine modification of certain protein related to apoptosis.Results The A549/DDP cells resisted the cytotoxic effect of cisplatin and the resistance index was 10.82.The inhibition of curcumin on A549 cells and A549/DDP cells was shown in the dose-dependent and time-dependent manner(P＜0.05)following 48 h treatment with curcumin.Value of curcumin to A549 cells was approximated to that of A549/DDP cells.Under fluorescence microscope,cells treated with curcumin showed shrunk or distorted nucleus with fluorescence conglomerated like grain in them.The apoptosis indexes of A549/DDP cells induced by curcumin were significantly different from those of the control.And the level of caspase-8 plO protein of A549/DDP cells induced by curcumin increased as the concentration of curcumin increased.Conclusions A549/DDP is sensitive to curcumin.Curcumin can inhibit A549/DDP cells from proliferating and induce cell apoptosis.The activation of caspase8 is one of the mechanisms.