全身和局部类固醇激素水平可能作为乳腺癌患者新的预后和预测生物标志物。我们旨在开发一种新的液相色谱-串联质谱(LC-MS/MS)方法,用于同时测量多个,人血清和乳腺癌组织中生物学关键的类固醇激素。
■定量方法由液-液萃取组成,SephadexLH-20色谱法用于组织提取物,并通过液相色谱-串联质谱法分析类固醇激素。我们分析了16和40例乳腺癌患者的血清和组织类固醇激素水平,分别,并评估其与临床参数的相关性。
■该方法包括量化血清中的九种类固醇激素[包括皮质醇,可的松,皮质酮,雌酮(E1),17β-雌二醇(E2),17α-羟孕酮,雄烯二酮(A4),睾酮和孕酮)和六个(包括可的松,皮质酮,E1,E2,A4和睾丸激素)在癌组织中。定量下限在血清(250μl)的0.003-10ng/ml和组织(20mg)的0.038-125pg/mg之间,分别。准确度在98%-126%之间,测定内变异系数(CV)低于15%,和试验间CV低于11%。组织的分析回收率在76%-110%之间。组织E1水平与组织E2水平呈正相关(p<0.001),血清E1、E2和A4水平(p<0.01)。组织E2水平与血清E1水平呈正相关(p=0.02),但与血清E2水平无关(p=0.12)。在肿瘤较大的患者中,组织E2的水平和E1与A4水平的比率(芳香化酶活性的指数)显着升高(分别为p=0.03和p=0.02)。
■该方法简便,适用于血清中临床重要的类固醇激素的特异性和准确的谱分析。然而,在组织样本中的类固醇分析中,概况方法的灵敏度是有限的,但如果样品的大小或其类固醇含量足够,则可用于分析乳腺癌组织中的类固醇。
UNASSIGNED: Systemic and local steroid hormone levels may function as novel prognostic and predictive biomarkers in breast cancer patients. We aimed at developing a novel liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the simultaneous measurement of multiple, biologically pivotal steroid hormones in human serum and breast cancer tissue.
UNASSIGNED: The quantitative method consisted of liquid-liquid extraction, Sephadex LH-20 chromatography for tissue extracts, and analysis of steroid hormones by liquid-chromatography-tandem mass spectrometry. We analyzed serum and tissue steroid hormone levels in 16 and 40 breast cancer patients, respectively, and assessed their correlations with clinical parameters.
UNASSIGNED: The method included quantification of nine steroid hormones in serum [including cortisol, cortisone, corticosterone, estrone (E1), 17β-estradiol (E2), 17α-hydroxyprogesterone, androstenedione (A4), testosterone and progesterone) and six (including cortisone, corticosterone, E1, E2, A4, and testosterone) in cancer tissue. The lower limits of quantification were between 0.003-10 ng/ml for serum (250 µl) and 0.038-125 pg/mg for tissue (20 mg), respectively. Accuracy was between 98%-126%, intra-assay coefficient of variations (CV) was below 15%, and inter-assay CV were below 11%. The analytical recoveries for tissue were between 76%-110%. Tissue levels of E1 were positively correlated with tissue E2 levels (p<0.001), and with serum levels of E1, E2 and A4 (p<0.01). Tissue E2 levels were positively associated with serum E1 levels (p=0.02), but not with serum E2 levels (p=0.12). The levels of tissue E2 and ratios of E1 to A4 levels (an index for aromatase activity) were significantly higher in patients with larger tumors (p=0.03 and p=0.02, respectively).
UNASSIGNED: The method was convenient and suitable for a specific and accurate profiling of clinically important steroid hormones in serum. However, the sensitivity of the profile method in steroid analysis in tissue samples is limited, but it can be used for the analysis of steroids in breast cancer tissues if the size of the sample or its steroid content is sufficient.