Rapid, high-resolution procedure for assessment of estrogen receptor heterogeneity in clinical samples.
Rapid, high-resolution procedure for assessment of estrogen receptor heterogeneity in clinical samples.
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用于评估临床样本中雌激素受体异质性的快速、高分辨率程序。
DOI：
10.1016/s0021-9673(01)81666-6
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发表时间：
1985
期刊：
影响因子：
--
通讯作者：
Wittliff,JL
中科院分区：
文献类型：
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作者：
Boyle,DM;Wiehle,RD;Shahabi,NA;Wittliff,JL
如果病灶含有合理水平的雌激素受体（> 10 fmol/mg 胞质蛋白），大约三分之二的乳腺癌女性可能会受益于激素治疗。我们的实验室和其他实验室表明，不仅受体的浓度，而且受体的各种分子形式可能是预测患者反应性的重要因素。到目前为止，这种异质性是通过分析需要 16 小时离心的蔗糖密度梯度来确定的。其他可以揭示受体亚型概况的方法因不可重复性、回收率差或进行分析所需的时间长度而受到损害。我们相信，阴离子交换和色谱聚焦模式下的高效液相色谱 (HPLC) 可能能够比目前任何其他方法更深入地了解受体结构。利用高活性配体 [16α-125I]碘雌二醇-17β (2200 Ci/mmol) 和流通设备监测电导率、pH 和放射性，我们能够描述乳腺癌子宫粗胞浆制剂中离子亚型的分布。这种“在线”技术快速（色谱图在 120 分钟内完成）、灵敏（明显相当于 1 fmol 的受体异构体）、高效（色谱柱通常返回 > 90% 的应用放射性标记受体）且可重复（由于现代 HPLC 设备的先进性）。很明显，这些技术可用于临床环境，以更好地描述雌激素受体亚型的概况，并且应该作为将受体结构与反应组织中激素功能相关联的方法进行探索。
Approximately two-third of the women with breast cancer may benefit from hormone therapies if the lesion contains estrogen receptor in reasonable levels (> 10 fmol/mg cytosol protein). Our lab and others have suggested that not only the concentration, but the various molecular form(s) of the receptor may be an important factor in predicting patient responsiveness. Until now, this heterogeneity has been determined by analysis on sucrose density gradients requiring 16 h of centrifugation. Other methodologies which can disclose the profile of receptor isoforms are compromised by irreproducibility, poor recovery, or the length of time required to perform the analysis. We believe that high-performance liquid chromatography (HPLC) in the anion-exchange and chromatofocusing modes may be able to supply more insight into receptor structure than currently available by any other method. Utilizing the high activity ligand [16α-125I]iodoestradiol-17β (2200 Ci/mmol) and flow-through equipment to monitor conductivity, pH, and radioactivity, we are able to describe the distribution of ionic isoforms in crude cytosolic preparations of breast cancer uterus. This “on-line” technology is rapid (chromatogram is complete within 120 min), sensitive (receptor isoforms equivalent to 1 fmol are apparent), efficient (columns typically return >90% of applied radio-labeled receptor) and reproducible (due to the sophistication of modern HPLC equipment). It is clear these techniques may be used in the clinical setting to better describe the profile of estrogen receptor isoforms and should be explored as a method of correlating receptor structure with hormone function in responsive tissues.