【摘要】 目的 研究人全长雄激素受体(androgen receptor,AR)cDNA质粒(pSAR-IRES-EGFP)在人进展期前列腺癌细胞系PC-3中稳转染再表达,即人前列腺癌细胞系PC-3-AR~+的肿瘤生物学特性及其干细胞相关基因蛋白表达的变化,探讨AR的分子生物学作用。方法(1)应用体外细胞培养,通过四氮甲基唑氮(MTT)和细胞划痕等方法检测癌细胞生长和迁移特性;Western Blot方法检测干细胞相关蛋白的表达;细胞爬片和免疫荧光方法检测干细胞相关蛋白的定位和表达。(2)应用人前列腺癌细胞组织重组-先天免疫缺陷小鼠(Nude)肾包膜下移植技术,建立人前列腺癌-小鼠移植瘤模型,通过H.E.染色和病理学分析研究癌细胞的成瘤性、浸润和转移能力等肿瘤生物学特性;通过免疫组化染色技术检测移植瘤干细胞相关蛋白的定位和表达。结果 人雄激素非依赖型进展期前列腺癌细胞系PC-3再表达全长AR.cDNA,即PC-3-AR+,导致以下情况的改变:(1)体外细胞培养中癌细胞的生长和扩增能力以及迁移特性均受抑制;(2)小鼠体内移植瘤肿瘤体积减小,癌细胞变大,分化增强,坏死增加和浸润能力减低;(3)培养细胞中检测到干细胞相关蛋白CD133、CD44和ALDH等的表达增加。结论 人雄激素非依赖型前列腺癌细胞系PC-3再表达全长AR cDNA,使其生长受抑,分化增强,坏死增加和干细胞相关基因蛋白(CD133、CD44和ALDH)的表达增加。更多还原

【Abstract】 Objective To investigate the changes of biological behaviors and sternness-related gene expression levels in human prostate cancer cells PC-3-AR~+ with androgen receptor(AR)-overexpressed and discuss its possible function.Methods(1) Cancer cell growth and migration were respectively measured by MTT assay and wound healing assay.The expressions of sternness-related proteins were detected by Western blot.The expression pattern and localization of sternness-related proteins were detected by cell immunofluerescence staining.(2) The human prostate cancer xenografting mouse models were established to detect the expression pattern and localization of sternness-related proteins and analyze histopathological changes.The models were established by implanting the tissue recombinants into the sub-renal capsule of severe combined immunodeficient(Nude) mice.Results Overexpression of full-length androgen receptor(AR) in human prostate cancer cell line PC-3-AR+ could lead to the following results.(1) The characteristics of cell growth and migration were inhibited;(2) The differentiation ability of xenografted cancer cells was increased,the tumor size and invasion ability were decreased.(3) The expression levels of sternness-related proteins,such as CD133,CD44 and ALDH,were enhanced.Conclusion The overexpression of full-length AR cDNA in human prostate cancer cells may result in the changes of cancer biological behaviors and sternness-related proteins expression.更多还原