文献类型：期刊文献

中文题名：DARPP-32基因对SH-SY5Y细胞药物敏感性的调节作用

英文题名：Effects of DARPP-32 on the drug sensitivity of SH-SY5Y cells

作者：张宏卫[1];刘丽华[2];李伟[3]

机构：[1]绍兴文理学院医学院附属医院神经科;[2]绍兴文理学院医学院分子诊断实验室;[3]四川大学华西医院神经科

年份：2007

卷号：23

期号：5

起止页码：977

中文期刊名：中国病理生理杂志

外文期刊名：Chinese Journal of Pathophysiology

收录：CSTPCD、、北大核心2004、CSCD2011_2012、北大核心、CSCD

语种：中文

中文关键词：神经母细胞瘤;基因;DARPP-32;RNA干扰;SH—SY5Y细胞

外文关键词：Neuroblastoma; Genes, DARPP-32; RNA interference; SH-SY5Y cells

中文摘要：目的:探讨DARPP-32基因对人神经母细胞瘤细胞药物敏感性的调节作用。方法:构建DARPP-32基因的真核表达载体和小干扰RNA载体,并将它们转导入人神经母细胞瘤细胞SH-SY5Y,G418筛选后获得稳定转染的阳性克隆后,应用RT-PCR和Western blotting进行鉴定;MTT法检测细胞转染前后药物敏感性的变化;流式细胞仪检测细胞转染前后对阿霉素的蓄积浓度的变化;RT-PCR和Western blotting检测细胞转染前后耐药相关蛋白P-gp、MRP和凋亡相关蛋白Bcl-2、Bax的表达变化。结果:成功构建了DARPP-32基因的真核表达载体和小干扰RNA载体;筛选到稳定的DARPP-32高/低表达的神经母细胞瘤细胞模型;MTT试验和流式细胞仪检测结果显示,上调DARPP-32的表达能够显著增强神经母细胞瘤细胞对长春新碱、阿霉素、5-氟尿嘧啶和顺铂的敏感性,提高细胞内阿霉素的蓄积(P<0.05),转染DARPP-32小干扰RNA后的细胞对化疗药物的敏感性降低,细胞内的阿霉素蓄积量显著减少(P<0.05);RT-PCR和Westernblotting结果显示,DARPP-32能够下调P-gp和Bcl-2的表达。结论:DARPP-32基因能够调节神经母细胞瘤细胞对化疗药物的敏感性,具有较好的临床应用前景。

外文摘要：AIM ： To investigate the effects of DARPP - 32 on the drug sensitivity of human neuroblastoma cell line SH -SYSY. METHODS： The plasmid containing cDNA of DARPP -32 gene and the small interfering RNA eukaryotic expression vector specific to human DARPP - 32 gene were constructed and transfected into SH - SYSY cells. Stable transfectants were identified by RT - PCR and Western blotting analysis. MTT assay and flow cytometry assay were used to investigate the effects Of DARPP - 32 on the drug sensitivity and the drug accumulation in cell models. The expressions of Bcl - 2, Bax, P - gp and MRP were analyzed by RT - PCR and Western blotting. RESULTS ： The stable clones with in- creased or decreased DARPP - 32 expression were successfully established. Up - regulation of DARPP - 32 significantly enhanced the drug sensitivity and the drug accumulation of SH - SYSY ceils due to down - regulation of P - gp and Bc1-2 expressions. Down - regulation of DARPP - 32 significantly reduced the drug sensitivity of SH - SYSY ceils and decreased drug accumulation in SH - SYSY ceils. CONCLUSION： DARPP - 32 might mediate the sensitivity of SH - SYSY ceils to conventional chemotherapeutic agents.