晚期糖基化终末产物受体胞外段不同功能段在前列腺癌细胞中的表达与定位

目的构建晚期糖基化终末产物受体(RAGE)胞外段不同功能段V/VC1真核细胞表达载体,并在前列腺癌细胞株PC-3中表达,为进一步研究其在前列腺癌发病中的作用和机制打下基础。方法采用PCR方法扩增RAGE不同功能段V/VC1的序列,利用分子克隆技术将其重组于pcDNA3-HA载体中。利用PCR和测序鉴定克隆正确性。转染PC-3细胞,用Western blotting检测其表达,免疫荧光检测其在细胞中的定位。结果克隆的RAGE不同功能段V/VC1真核表达载体完全正确,Western blotting检测到RAGE不同功能段V/VC1的表达,免疫荧光检测其主要定位于细胞浆中。结论成功地构建了RAGE不同功能段V/VC1真核表达载体,其能够在PC-3细胞中表达。

Construction of eukaryotic expression vectors for different domains of the extracellular region of RAGE and their expression in prostate cancer cells

Objective To construct eukaryotic expression vectors for different domains(V and VC1) of the extracellular region of the receptor of advanced glycation end products(RAGE) and investigate the roles of these domains in prostate cancer.Methods The coding sequence of V and VC1 domains was amplified from the plasmid pcDNA3-HA-RAGE by PCR and cloned into the pcDNA3-HA vector following routine procedures.After identification by PCR and sequencing,the vectors including V and VC1 domains were transfected into PC-3 cells.Western blotting and immunofluorescence were used to detect the expression and distribution of the expressed products in transfected PC-3 cells.Results The expression vectors containing V and VC1 domains of RAGE were successfully constructed as confirmed by PCR and DNA sequence analysis.The V and VC1 domains of RAGE were highly expressed and showed a cytoplasmic distribution in transfected PC-3 cells.Conclusion The constructed eukaryotic expression vectors for V and VC1 domains of RAGE can be efficiently expressed in prostate cancer cells.