氟他胺联合去雄激素环境诱导的雄激素非依赖性前列腺癌细胞模型的建立

Establishment of an Androgen-independent Prostate Cancer Cell Line Induced by the Combined of Flutamide and Androgen-free Environment

采用氟他胺联合去雄激素环境联合诱导培养的方法在体外建立一株雄激素非依赖性前列腺癌细胞系。倒置显微镜下观察细胞形态学变化;采用CCK-8法测定细胞生长率;流式细胞技术检测细胞周期和Bcl-2、Bax的表达变化。结果显示,第1～20代实验组细胞形态瘦长,第30代后细胞大部分呈扁平状,同时呈聚集生长,而对照组细胞形态始终呈长梭形;不同代数的实验组LNCaP细胞在氟他胺浓度为5.0×10-6mol/L的去雄激素培养液中的细胞生长率总体呈上升趋势;氟他胺和去雄激素环境对实验组LNCaP细胞的细胞周期均无明显影响,而对照组LNCaP细胞的生长受氟他胺和去雄激素环境影响而受到抑制;实验组第60代LNCaP细胞Bcl-2平均荧光强度明显高于对照组,而Bax平均荧光强度与对照组间无显著差异。结果表明LNCaP细胞在氟他胺和去雄激素环境下能形成雄激素非依赖性前列腺癌细胞,这为其发病机制的研究提供了合适的实验对象。

We established an androgen-independent prostate cancer cell line induced by the combination of flutamide and androgen-free environment after long-term culture in vivo. Cell morphological changes were traced with inverted microscope. Cell proliferation rate was measured by CCK-8 assa. Flow cytometry was used to determine the changes of cell cycle, and expression of Bcl-2 and Bax. Cell morphology of experimental group from i to 20 passages was leptosomic, however, it was changed into flatting and aggregation growth pattern after 30 passages, cell morphology of control group was always shuttle-like shape. Cell growth rate of experimental group LNCaP cell was shown a gradual uptrend in the whole process when cultured in androgen-free medium with the terminal concentration of flutamide of 5.0×10^-6 mol/L. Flutamide and androgen-free environment was revealed to have no significant effect on the cell cycle of experimental group LNCaP cell, but the control group was found to be inhibited. Mean fluorescence intensity （MFI） of Bcl-2 in experimental group was significantly higher than control group. However, there was no significant difference in the MFI of Bax between two groups. LNCaP cell could be changed into androgen-independent prostate cancer cell after long-term induced by the combination of flutamide and androgen-free environment, and this would be a suitable cell line for the pathogenesis studies.