羟基磷灰石法和葡聚糖活性炭法测定雌激素受体的比较

A comparison between HAP and DCC methods for estrogen receptor measurements.

目的：建立一种比葡聚糖活性炭（ＤＣＣ）法更灵敏、准确、简便的羟基磷灰石（ＨＡＰ）法测定雌激素受体（ＥＲ）的含量。方法：以ＳＤ雌性大鼠子宫胞浆抽提液为材料，用ＨＡＰ法多点饱和分析和单点饱和分析ＥＲ含量；比较在测定反应中总蛋白量对ＨＡＰ法和ＤＣＣ法测定值的影响，并比较两者在３０例乳腺癌和３０例子宫肌瘤中ＥＲ测定值的差异。结果：大鼠子宫胞浆ＥＲ含量为１３５ｆｍｏｌ／ｍｇ蛋白，Ｋｄ＝０．５８ｎＭ。当测定反应中总蛋白量低于２００μｇ时，ＤＣＣ法测定值严重失真，而ＨＡＰ法在总蛋白量为５０μｇ时仍能准确测出真实值。在３０例乳腺癌和３０例子宫肌瘤ＥＲ测定值中，当ＥＲ含量较低时，ＤＣＣ法测定值偏低甚至有假阴性．而ＥＲ含量高时两者接近。在ＥＲ含量较高时，两者测定值相近。结论：ＨＡＰ法比ＤＣＣ法灵敏、可靠。用以测定甾体激素高非特异性结合的靶组织与非靶组织中的ＥＲ低含量时，ＨＡＰ法更有价值。

Objective To establish HAP assay a powerful method to determine estrogen receptor more sensitively,more accurately and more conveniently than that of DCC assay. Methods Uterine cytosol extracts from SD female rats were used to establish the HAP multi point saturation assay and HAP single point saturation assay.The authors examined the effect of total protein level on HAP and DCC assays and compared the ER values measured by HAP and DCC in 30 breast cancer cases and 30 leiomyoma cases respectively. Results The ER value in rat uterine cytosol extract was 135 fmol/mg cytosol protein.Kd value was 0.58nM.HAP assay could exactly determine ER value when total protein was less than 50μg,but DCC assay could not determine even when total protein level was less than 200μg.When ER level in 30 breast cancer cases and 30 leiomyoma cases was low,the value examined by DCC assay was lower than its real value,but when ER level was high enough,the value measured by DCC and HAP were almost equal to each other. Conclusions HAP assay is a more sensitive,more accurate method than that of DCC assay and plays a more valuable role on determinating low ER value in all kinds of steroid target and nontarget tumor tissues with high nonspecific binding.