摘要
为了获得代表不同前列腺癌进展阶段的细胞系的胞外蛋白表达谱,验证其中差异表达蛋白是否为分泌蛋白,在细胞水平看其是否有作为前列腺癌血清标志物的潜质,文章利用双向电泳寻找胞外蛋白中差异表达的点,并质谱鉴定其是何种蛋白质。应用RT-PCR方法分析候选分子在8种细胞系中的表达和对雄激素刺激的应答,构建了候选分子的真核表达载体,瞬时转染293T细胞,应用标签抗体Western blotting方法检测验证细胞培养基中候选分子的表达。结果表明:筛选出两个C4-2胞外高表达的分子--磷酸丙糖异构酶-1(Triosephosphate isomerase1,TPI1)和多配体聚糖结合蛋白(Syndecan binding protein,syntenin,ST1);转录水平发现它们与前列腺癌恶性程度相关,并且后者受雄激素的作用下调;二者均为分泌蛋白。磷酸丙糖异构酶-1和多配体聚糖结合蛋白均有作为指示前列腺癌发展阶段的血清标志物的潜质。
Our research intends to obtain extra-cellular proteinogram of cell lines representing different advancement stages of prostate cancer and to test whether screened differential expression proteins can be secreted and used as serum biomarkers for prostate cancer.By examining differential expression spots in two extra-cellular protein 2D-PAGE gels and mass spectrum,candidate molecules were obtained.The expressions of these candidate molecules in eight cell lines and response to androgen stimulus in LNCaP were analyzed by RT-PCR.By constructing eukaryotic expression vectors and western-blotting with anti tags antibodies,the candidate molecules were tested to understand whether they can be expressed in tansfected 293T cell culture fluid.Two overexpressed molecules-triosephosphate isomerase 1 （TPI1） and syndecan binding protein,syntenin （ST1）-in extra-cellular proteinogram of C4-2 were screened out;both of them are secretary proteins.On transcriptional level,both proteins were up-regulated with the malignancy of prostate cancer cell lines and ST1 was dose-dependently inhibited by androgen.Considering cellular level results,both TPI1 and ST1 have their potential as serum biomarkers for indicating the developmental stage of prostate cancer.
出处
《遗传》
CAS
CSCD
北大核心
2010年第3期235-241,共7页
Hereditas（Beijing)
基金
国家高技术研究发展计划项目(编号:2008AA02Z123)
国家自然科学基金项目(编号:30770834
30870961)资助
