EGF与miRNA-21联合构建雄激素非依赖性前列腺癌LNCaP亚细胞模型  

Establishment of androgen-independent human prostate cancer sub line cell model of LNCaP with EGF and miRNA-21

作　　者：李刚[1] 佟明[1] 金艳阳[1] 刘四明[1] 

机构地区：[1]辽宁医学院附属第一医院,辽宁锦州121001

出　　处：《山东医药》2010年第52期32-34,共3页Shandong Medical Journal

摘　　要：目的构建雄激素非依赖性的人前列腺癌细胞株LNCaP亚型。方法前期常规RPMI 1640培养基培养LNCaP细胞,然后给予无酚红的RPMI 1640培养基,其中加入10%活性碳/葡聚糖处理的胎牛血清,细胞传代后将10μg/L表皮生长因子(EGF)加入培养基,50 nmol/L miRNA-21类似物转染细胞,长期培养。镜下观察细胞形态变化,MTT检测细胞增殖活性,RT-PCR检测雄激素受体基因扩增情况。结果细胞培养约2.5个月后,LNCaP细胞成功转变为非雄激素依赖的前列腺癌细胞LNCaP亚细胞,细胞增殖能力升高,雄激素受体表达上调。结论体内雄激素依赖性前列腺癌可向雄激素非依赖性前列腺癌转变,而且miRNA-21和EGF对该转变起促进作用。Objective To establish an androgen-independent human prostate cancer cell sub line of LNCaP.Methods LNCaP cells were cultured in conventional RPMI 1640 medium early,then were changed to phenol red-free RPMI 1640 medium in which there was 10% fetal bovine serum treated by carbon and dextran.After passage,10 μg/L epidermal growth factor（EGF） was kept into medium and transfected cell with 50 nmol/L miRNA-21 analog.The cells were cultured for long time.To observe the changes of cell morphology through microscopy,and to detecte the power of cell proliferation and test the amplification of androgen receptor（AR） gene with RT-PCR method.Results After two and a half months,LNCaP cells transformed into androgen-independent prostate cancer LNCaP sub line cells successfully.Cell proliferation increased.The expression of AR was up regulation.Conclusions The androgen-dependent prostate cancer cells can transform into androgen-independent prostate cancer cells.EGF and miRNA-21 play an improving role in this change.

关 键 词：前列腺肿瘤 雄激素非依赖性 MIRNA-21 表皮生长因子 LNCAP 

分 类 号：R737.25[医药卫生—肿瘤]