CAJ | 학술논문

目的观察17-二甲胺乙胺基-17-去甲氧基格尔德霉素（17-DMAG）对人结肠癌HT-29细胞增殖、细胞凋亡的影响及其细胞内活性氧（ ROS）的变化。方法用CCK-8检测17-DMAG对结肠癌HT-29细胞增殖影响；AnnexinV-FITC/PI双染法流式细胞检测细胞凋亡率；酶标仪检测细胞内ROS的变化。结果17-DMAG呈时间-剂量依赖性抑制HT-29细胞增殖。0.25μmol/L、0.5μmol/L、1.0μmol/L、2.5μmol/L作用于HT-29细胞24 h后，细胞增殖抑制率分别为（22.17±1.15）％、（28.45±1.16）％、（35.04±1.58）％和（46.85±2.44）％，作用48 h后，细胞增殖抑制率分别为（27.55±0.65）％、（33.33±1.23）％、（46.20±4.76）％和（55.45±4.47）％，作用72 h，细胞增殖抑制率分别为（39.19±1.74）％、（44.29±2.00）％、（50.66±2.17）％和（58.84±3.18）％。正常对照组 HT-29细胞24 h 的自然总凋亡率（早期＋晚期）为（2.72±0.57）％，0.25μmol/L、0.5μmol/L、1.0μmol/L和2.5μmol/L 17-DMAG干预24 h后细胞总凋亡率分别为（5.38±0.46）％、（6.88±0.52）％、（10.44±0.32）％与（17.87±4.66）％，17-DMAG作用HT-29细胞24 h的凋亡率与对照组细胞凋亡率相比差异有统计学意义（ P ＜0.05）。0.25μmol/L、0.5μmol/L、1.0μmol/L 和2.5μmol/L 17-DMAG作用HT-29细胞12 h与24 h，其活性氧水平均较对照组有不同程度的上升，差异有统计学意义（ P＜0.05）。结论17-DMAG体外呈时间-剂量依赖性抑制HT-29细胞增殖，诱导其凋亡，可能部分与细胞内的ROS升高有关。
목적관찰17-이갑알을알기-17-거갑양기격이덕매소（17-DMAG）대인결장암HT-29세포증식、세포조망적영향급기세포내활성양（ ROS）적변화。방법용CCK-8검측17-DMAG대결장암HT-29세포증식영향；AnnexinV-FITC/PI쌍염법류식세포검측세포조망솔；매표의검측세포내ROS적변화。결과17-DMAG정시간-제량의뢰성억제HT-29세포증식。0.25μmol/L、0.5μmol/L、1.0μmol/L、2.5μmol/L작용우HT-29세포24 h후，세포증식억제솔분별위（22.17±1.15）％、（28.45±1.16）％、（35.04±1.58）％화（46.85±2.44）％，작용48 h후，세포증식억제솔분별위（27.55±0.65）％、（33.33±1.23）％、（46.20±4.76）％화（55.45±4.47）％，작용72 h，세포증식억제솔분별위（39.19±1.74）％、（44.29±2.00）％、（50.66±2.17）％화（58.84±3.18）％。정상대조조 HT-29세포24 h 적자연총조망솔（조기＋만기）위（2.72±0.57）％，0.25μmol/L、0.5μmol/L、1.0μmol/L화2.5μmol/L 17-DMAG간예24 h후세포총조망솔분별위（5.38±0.46）％、（6.88±0.52）％、（10.44±0.32）％여（17.87±4.66）％，17-DMAG작용HT-29세포24 h적조망솔여대조조세포조망솔상비차이유통계학의의（ P ＜0.05）。0.25μmol/L、0.5μmol/L、1.0μmol/L 화2.5μmol/L 17-DMAG작용HT-29세포12 h여24 h，기활성양수평균교대조조유불동정도적상승，차이유통계학의의（ P＜0.05）。결론17-DMAG체외정시간-제량의뢰성억제HT-29세포증식，유도기조망，가능부분여세포내적ROS승고유관。
Objective To investigate the effects of HSP 90 inhibitor 17-DMAG on proliferation and apoptosis of colon cancer HT-29 cells and the intracellular level of reactive oxygen species .Methods HT-29 cells were treated with 17-DMAG.The cell proliferation inhibition rate was evaluated by CCK-8 assay.Annexin V PI double labeling FCM was used to determine cell apoptotic rate .Intracellular reactive oxygen species ( ROS) generation was measured by microplate reader .Results 17-DMAG time-dose-dependently inhibit the proliferation of HT-29 cells, after 0.25 μmol/L, 0.5 μmol/L, 1.0 μmol/L and 2.5 μmol/L 17-DMAG exposure for 24 hours, the cell proliferation inhibition rate was ( 28.45 ±1.16 )%, ( 35.04 ±1.58 )%, ( 46.85 ±2.44 )% respectively , after exposure for 48 hrs,the cell proliferation inhibition rate was increased to (27.55 ±0.65)%,(33.33 ±1.23)%, (46.20 ±4.76)%,(55.45 ±4.47)% respectively,after exposure for 72 hours,the cell proliferation inhibition rate was to(39.19 ±1.74)%,(44.29 ±2.00)%,(50.66 ±2.17)%,(58.84 ±3.18)%;17-DMAG can markedly promote apoptosis ,when HT-29 cells were exposed to 0.25 μmol/L,0.5 μmol/L,1.0 μmol/L and 2.5 μmol/L 17-DMAG for 24 hours,the total apoptotic rate for 24 hours was increased to (5.38 ±0.46)%,(6.88 ±0.52)%, (10.44 ±0.32)% and(17.87 ±4.66)% respectively,compared to(2.72 ±0.57)% HT-29 cells without 17-DMAG( P<0.05 ) .Compared to the control group ,the level of intracellular ROS in the 17-DMAG-treated group for 12 hours or 24 hours was relatively higher ( P<0.05 ) .Conclusions 17-DMAG can time-dose-dependently inhibit proliferation and induce apoptosis of HT-29 cells,and this effect may be partly associated with an intracellular ROS increase .