摘要
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24h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48h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72h,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24h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/L17-DMAG干预24h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24h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/L17-DMAG作用HT-29细胞12h与24h,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。
Objective To investigate the effects of HSP90 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 μ mol/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 ceils, and this effect may be partly associated with an intracellular ROS increase.
出处
《中华临床医师杂志（电子版）》
CAS
2013年第8期113-115,共3页
Chinese Journal of Clinicians(Electronic Edition)
基金
上海市公共卫生临床中心人才引进基金(RCJJP8)
