| 摘要: |
| 目的:利用激素递减法对LNCaP细胞进行去雄环境培养,建立雄激素非依赖的LNCaP细胞(LNCaP-AI)亚系并予以鉴定。方法:利用活性炭处理的血清培养模拟去雄细胞环境,逐步递减培养基中激素10 d,后完全在非雄培养,共连续培养3个月,直到LNCaP细胞重新进入快速生长期,利用CCK-8法、放免法、RT-PCR法分别测定细胞生长曲线、PSA及雄激素(AR)表达情况。结果:LNCaP细胞在激素递减后,生长缓慢,呈神经内分泌样改变和成簇生长,经过共3个月的连续非雄培养,细胞重新恢复以前的形态及生长。CCK-8测定提示LNCaP-AI细胞能够在去雄环境中生长,放免法提示LNCaP-AI细胞能够在去雄环境中恢复分泌PSA功能,RT-PCR方法提示LNCaP-AI细胞显著高表达AR。结论:激素递减方法可以3个月左右有效地建立雄激素非依赖的LNCaP细胞亚系。 |
| 关键词: 前列腺肿瘤 雄激素非依赖 LNCaP细胞系 |
| DOI：10.3724/SP.J.1008.2008.01311 |
| 投稿时间：2008-04-22修订日期：2008-09-10 |
| 基金项目: |
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| Establishment of androgen-independent human prostate cancer line LNCaP by gradual deprivation of hormone |
| JI Jia-tao,XU Chuan-liang,YE Hua-mao,ZHOU Tie,TANG Yuan-jie,SUN Ying-hao* |
| (Department of Urology,Changhai Hospital,Second Military Medical University,Shanghai 200433,China) |
| Abstract: |
| Objective:To establish and identify androgen-independent human prostate cancer cell line LNCap by culturing LNCaP cells with gradual deprivation of hormone.Methods: LNCaP cells were cultured in the medium with gradual deprivation of hormone (treated by active carbon to simulate androgen deprivation)for 10 days; and then the cells were cultured with complete deprivation of androgen for 3 months till the cell entered the rapid proliferation phase again.The cell growth and expression of PSA and androgen were examined by CCK-8,immunfluorescence and RT-PCR methods.Results: LNCaP cells grew slowly after deprivation of hormone and took on a neuroendocrine phenotype and cluster growth pattern.After 3 months’ non-androgen culture,the cells regained original morphology and growth.CCK-8 indicated that LNCaP cells could grow in non-androgen condition; immunofluorescence assay indicated that LNCaP-AI cells could regain PSA-secreting activity in non-androgen condition; and RT-PCR suggested that androgen was highly expressed in LNCaP-AI cells.Conclusion: Androgen-independent LNCaP cell line can be established by culturing with gradual deprivation of hormone for 3 months. |
| Key words: prostatic neoplasms androgen-independent LNCaP cell line |
