CAJ | 학술논문

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 isomerase 1,TPI1)和多配体聚糖结合蛋白(Syndecan bindingprotein,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.
위료획득대표불동전렬선암진전계단적세포계적포외단백표체보,험증기중차이표체단백시부위분비단백,재세포수평간기시부유작위전렬선암혈청표지물적잠질,문장이용쌍향전영심조포외단백중차이표체적점,병질보감정기시하충단백질.응용RT-PCR방법분석후선분자재8충세포계중적표체화대웅격소자격적응답,구건료후선분자적진핵표체재체,순시전염293T세포,응용표첨항체Western blotting방법검측험증세포배양기중후선분자적표체.결과표명:사선출량개C4-2포외고표체적분자--린산병당이구매-1(Triosephosphate isomerase 1,TPI1)화다배체취당결합단백(Syndecan bindingprotein,syntenin,ST1);전록수평발현타문여전렬선암악성정도상관,병차후자수웅격소적작용하조;이자균위분비단백.린산병당이구매-1화다배체취당결합단백균유작위지시전렬선암발전계단적혈청표지물적잠질.
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 (TPI 1) 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.