Boyle D M, Wiehle R D, Shahabi N A, Wittliff J L
J Chromatogr. 1985 Jun 26;327:369-76. doi: 10.1016/s0021-9673(01)81666-6.
Approximately two-third of the women with breast cancer may benefit from hormone therapies if the lesion contains estrogen receptor in reasonable levels (greater than 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 [16 alpha-125I]iodoestradiol-17 beta (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 and 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 greater than 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.
如果乳腺病变组织中雌激素受体水平合理（大于10 fmol/mg胞浆蛋白），大约三分之二的乳腺癌女性患者可能从激素疗法中获益。我们实验室以及其他一些机构已表明，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一个重要因素。到目前为止，这种异质性是通过在蔗糖密度梯度上进行分析来确定的，该分析需要16小时的离心。其他能够揭示受体亚型谱的方法存在不可重复性、回收率低或分析所需时间长等问题。我们认为，阴离子交换和色谱聚焦模式的高效液相色谱（HPLC）可能比目前任何其他方法都能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）以及用于监测电导率、pH值和放射性的流通式设备，我们能够描述乳腺癌和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（相当于1 fmol的受体亚型清晰可见）、高效（柱通常能返回大于90%的应用放射性标记受体）且可重复（由于现代HPLC设备的精密性）。很明显，这些技术可用于临床，以更好地描述雌激素受体亚型谱，并且应该作为一种将受体结构与反应性组织中的激素功能相关联的方法进行探索。
