CAJ | 학술논문

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.
목적 연구강황소대인폐선암A549/DDP세포증식적억제작용급기대세포조망적영향.방법 채용사갑기우담서람(MTT)측정세포대순박적민감성화강황소대세포증식적억제작용;도치상차현미경화형광현미경하관찰세포형태학적변화;응용원위말단표기(TUNEL)염색급류식세포의검측세포조망정황;이Western blot분석조망상관단백표체변화.결과 실험중소용적A549/DDP세포대순박내약,내약지수위10.82.강황소대A549급A549/DDP세포증식균유억제작용,병차정시간、농도의뢰성(P＜0.05),대량세포주적반수억제농도분별위16.28/μmol/L화18.06μmol/L,차이무통계학의의(P＞0.05).강황소처리A549/DDP세포후,세포변형、축소、탈락.Hoechst염색현시A549/DDP세포핵축소、변형,치밀농염,형광성단괴분포.TUNEL법이급류식세포의분석결과시세포조망솔정명현제량의뢰관계(P＜0.05).Western blot결과현시,수강황소농도적증가,천동안산특이성반광안산단백매8(cysteinyl aspartate-specific protease-8,caspase-8)p10단백수평증가.결론 A549/DDP세포대강황소무항약성,강황소대인폐선암A549/DDP세포증식구유억제작용,병능유도세포조망.caspase-8적활화시기중적궤제지일.
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.