摘要
目的探讨熊去氧胆酸(UDCA)对肝癌细胞株HepG2增殖、凋亡及Survivin、Caspase-3表达的影响。方法UDCA孵育细胞后,分别采用MTT法、流式细胞仪、激光共聚焦显微镜、半定量RT-PCR法等观测相关指标。结果UDCA能抑制HepG2增殖,诱导其凋亡,并具有浓度、时间依赖性。随着Survivin表达下调,Caspase-3表达上调(r=-0.9959,P<0.01)、细胞增殖抑制(r=0.9883,P<0.05)、凋亡指数增加(r=-0.9521,P<0.05)。结论UDCA可能通过干预Survivin的表达,减弱后者对Caspase-3的负调控作用,而影响肝癌细胞的增殖、凋亡。
Objective to investigate the effects of UDCA on the growth and expression of Survivin, Caspase-3 of human hepatoma carcinoma （HCC） cellline HepG2, and to discuss the possible mechanism. Methods I-ICC cell line HepG2 was treated by UDCA in 12.5, 25, 50, 100,200,400μg/ml respectively, and was detected at 6, 12, 18,24,48, 72h by MTT assay, flow cytometry （FCM） with PI staining or Annexin V-FITC/PI double stainings, optical microscope,laser scanning confocal mieroscopy, semiquantitative RT-PCR. Results Cell proliferation was blocked by UDCA, which showed significant positive association with the concentration and the duration of UDCA treatment（6h, r = 0.9775, P 〈 0.01 ; 12h, r = 0. 8743, P 〈0.05; 24h, r=0.9259, P〈0.01; 36h, r=0.9499, P〈0.01; 72h, r=0.8352, P 〈0.05）. Optical microscope showed that the amount of round, small and dead cells increased after treated by UDCA. Laser confocal microscopy also showed the presence of apoptosis of cells after the treatment. FCM analysis showed that the apoptotic effect of UDCA on cell line HepG2 was concentration-and time-dependent. On the concentration of 200 mg/ml ,it was after 6h that apoptosis rate, （ 12.17 ＋ 1.76） % ,began to rise,singnificantly higher than that of control group, （7.92 ＋ 1.15） % . And at 24h, apoptosis rate of treatment group was higher than that of control group was P 〈 0.01. After 24 hours treated by UACA, on the concentration of 200ug/ml, the ratio of S-phase of control group was （48.09 ＋ 2.31） % , higher than that of control group （21.83 ＋ 2.21 ）%, P 〈 0.01. The longer UDCA treated, the less Survivin mRNA expressed , and the more that of Caspase-3 expressed. There was correlation between the expression of Survivin with Caspase-3 （ r = - 0. 9959, P 〈 0.01 ）, cellular proliferation （ r = 0. 9883, P 〈 0.05）, or apoptosis index （ r = - 0. 9521, P 〈 0.05） in our study. Conclusion The mechanism of UDCA inhibiting cellular proliferation and inducing apoptosis of HCC line HepG2 may possibly relate to influencing the expression of Survivin which inhibits the activity of Caspase-3.
出处
《胃肠病学和肝病学杂志》
CAS
2006年第5期492-496,共5页
Chinese Journal of Gastroenterology and Hepatology
作者简介
通讯作者：杨大明，E-mail：zzb0708@126．com
