论文部分内容阅读
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。
Objective To study the expression of human full-length androgen receptor (AR) cDNA plasmid (pSAR-IRES-EGFP) in human prostate cancer cell line PC-3. 3-AR ~ + tumor biological characteristics and changes in stem cell-related gene expression, explore the molecular biology of AR. Methods (1) In vitro cell culture was used to detect the growth and migration characteristics of cancer cells by MTT assay and cell scratch assay. The expression of stem cell related proteins was detected by Western Blot method. Cell proliferation and immunofluorescence The localization and expression of stem cell related proteins were examined. (2) The human prostate cancer-mouse xenograft model was established by subrenal transplantation of human prostate cancer cell-tissue-innate immunity-deficient mice (Nude). The tumorigenesis of cancer cells was studied by HE staining and pathological analysis Tumorigenicity, invasion, metastasis and so on. The localization and expression of tumor stem cell-related proteins were detected by immunohistochemical staining. Results The re-expression of full-length AR.cDNA, PC-3-AR +, in human androgen-independent advanced prostate cancer cell line PC-3 led to the following changes: (1) growth and expansion of cancer cells in vitro (2) The tumor volume of the transplanted tumor in mice decreased, the cancer cells became larger, the differentiation increased, the necrosis increased and the infiltration capacity decreased. (3) The stem cell-associated protein CD133 was detected in cultured cells, CD44 and ALDH expression increased. CONCLUSION: The full-length AR cDNA of human androgen-independent prostate cancer cell line PC-3 re-expresses and inhibits its growth, differentiation, enhancement of necrosis and increase of the expressions of stem cell related proteins (CD133, CD44 and ALDH).