芬太尼对人肝癌细胞bel-7404生长与凋亡的影响

Effect of fentanyl on growth and apoptosis of human hepatoma cell line bel-7404

目的探讨芬太尼对人肝癌细胞bel-7404生长及凋亡的影响。方法体外培养人肝癌bel-7404细胞,分F1、F2、F3、F4,4个实验组和1个对照组,实验组分别在RPMI-1640培养液中加入5ng/ml(F1)、50ng/ml(F2)、500ng/ml(F3)、5000ng/ml(F4)芬太尼,对照组不加芬太尼,所有样本孵育24h后,用倒置显微镜观察细胞形态学改变,应用MTT法和克隆形成实验检测细胞的增殖活性,应用流式细胞仪检测细胞凋亡率与细胞周期。结果各实验组人肝癌bel-7404细胞MTT和克隆形成率明显低于对照组(P<0.01),细胞凋亡百分比显著高于对照组(P<0.05)。芬太尼浓度≥50ng/ml时,随着浓度的增加,凋亡率逐渐增高,细胞周期中G0/G1期比例逐渐增加,S期细胞比例逐渐减少,与对照组比较差异具有统计学意义(P<0.05)。结论芬太尼可剂量依赖性抑制人肝癌细胞bel-7404的生长,干扰肝癌细胞增殖周期,诱导肝癌细胞凋亡。

Objectives To study the effects of fentanyl on growth and apoptosis of human hepatoma cell line bel-7404. Methods bel-7404 cells were. divided into four test groups （ F1, F2, F3 and F4 ） and a control group （C）. Cells in the four test groups were incubated in RPMI-1640 medium that contained different concentration of fentanyl respectively, such as 5,50,500 and 5000 ng/ml. Cells in the control group were cultured in RPMI-1640 medium only. After 24-hour incubation, morphological changes of the cells were observed by inverted microscope, the level of cell proliferation were evaluated by the methods of MTT and colony formation, and the distribution of cell cycle and the apoptosis rate were detected with flow cytometry （ FCM ）. Results The results from MTT and colony formation on the test groups were significantly lower than that of the control group （P 〈 0. 01 ）. The percentage of apoptosis was significantly higher in the test groups than that of the control group （P 〈 0.05 ）. When the concentration of fentanyl was ≥50ng/ml, the rate of apoptotic cells increased gradually with the increasing concentra- tion of fentanyl,and the ratio of cells in G0/G1 phase enhanced gradually and in S phase decreased gradually ,which was signifi- cantly different from that of the control group （P 〈 0. 05）. Conclusion The inhibitory effect of fentanyl on growth of bel-7404 cell is dose-dependent. Fentanyl is able to block cell cycle and to induce apoptosis.