A novel HPLC-RIA method for the simultaneous detection of estrone, estradiol and estrone sulphate levels in breast cancer tissue
A novel HPLC-RIA method for the simultaneous detection of estrone, estradiol and estrone sulphate levels in breast cancer tissue
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DOI：
10.1016/s0960-0760(00)00036-4
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发表时间：
2000-04-01
影响因子：
4.1
通讯作者：
Lonning, PE
中科院分区：
文献类型：
--
作者：
Geisler, J;Berntsen, H;Lonning, PE
剥夺雌激素是治疗激素敏感性乳腺癌的有效方法。虽然人们对绝经前女性去势和绝经后女性芳香酶抑制剂使用相关的血浆雌激素水平了解很多，但目前人们对肿瘤内雌激素产生的兴趣日益浓厚。然而，关于肿瘤内雌激素水平变化的知识是有限的，这主要是由于组织样本中雌激素分数测量的方法学问题。在这里，我们描述了一种同时测量乳腺肿瘤组织中三种主要雌激素成分：雌酮（E-1）、雌二醇（E-2）和硫酸雌酮（EIS）的新方法。与[H-3]标记的雌激素标准品一起孵育后，通过乙醚萃取分离粗级分。 E1S 级分用硫酸酯酶水解，然后在 Sephadex 柱上洗脱。在 RIA 分析之前，使用高压液相色谱 (HPLC) 纯化各个雌激素组分。将雌酮和E1S转化为E-2，最后使用雌二醇-6-(O-羧甲基)-肟基-2-(2-[I-125]-碘-组胺)作为配体，通过相同的高灵敏度和特异性放射免疫测定法测量所有三种雌激素组分。尽管使用了多个纯化步骤，但发现氚化雌激素的内部回收率为 E-1 和 E-2 的 25-50%，以及 E1S 的 15-30%。该方法的检测限分别为：E-2 为 4.3 fmol/g 组织、E-1 为 19.8 fmol/g 组织、E1S 为 11.9 fmol/g 组织。使用局部晚期乳腺癌组织 (n = 14)，我们发现 E-1、E-2 和 E1S 的中位水平分别为 283.8 fmol/g 组织（范围 19.8-547.5）、554.1 fmol/g（9.5-3024.2）和 209.4 fmol/g（11.9-753.4）。这里描述的方法是研究乳腺组织活检中肿瘤内雌激素分数的有前途的工具。 (C) 2000 Elsevier Science Ltd. 保留所有权利。
Estrogen deprivation is an effective approach for treatment of hormone sensitive breast cancer. While much is known about plasma estrogen levels with respect to castration in premenopausal women and use of aromatase inhibitors in postmenopausal women, currently there is increasing interest in intra-tumour estrogen production. However, knowledge about alterations in intra-tumour estrogen levels is limited, mainly due to methodological problems with measurements of estrogen fractions in tissue samples. Here we describe a new method for simultaneous measurement of the three main estrogen fractions, estrone (E-1), estradiol (E-2) and estrone sulphate (EIS) in breast tumour tissue. Following incubation with [H-3]-labelled estrogen standards, crude fractions were separated by ether extraction. The E1S fraction was hydrolysed with sulphatase followed by eluation on a Sephadex column. High pressure liquid chromatography (HPLC) was used to purify the individual estrogen fractions prior to RIA analysis. Estrone and E1S were converted into E-2, and all three estrogen fractions were finally measured by the same highly sensitive and specific radioimmunoassay using estradiol-6-(O-carboxymethyl)-oximino-2-(2-[I-125]-iodo-histamine) as a ligand. Although several purification steps were used, the internal recovery values for tritiated estrogens were found to be 25-50% for E-1 and E-2 and 15-30% for E1S. The detection limit of this method was 4.3 fmol/g tissue for E-2, 19.8 fmol/g tissue for E-1 and 11.9 fmol/g E1S, respectively. Using tissue from locally advanced breast cancers (n = 14), we found median levels of E-1, E-2 and E1S to be 283.8 fmol/g tissue (range 19.8-547.5), 554.1 fmol/g (9.5-3024.2) and 209.4 fmol/g (11.9-753.4), respectively. The method described here is a promising tool to study intra-tumour estrogen fractions in breast tissue biopsies. (C) 2000 Elsevier Science Ltd. All rights reserved.