Evaluation of a Luciferase-Based Reporter Assay as a Screen for Inhibitors of Estrogen-ERα-Induced Proliferation of Breast Cancer Cells
Evaluation of a Luciferase-Based Reporter Assay as a Screen for Inhibitors of Estrogen-ERα-Induced Proliferation of Breast Cancer Cells
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DOI：
10.1177/1087057112442960
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发表时间：
2012-08-01
影响因子：
--
通讯作者：
Shapiro, David J.
中科院分区：
文献类型：
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作者：
Andruska, Neal;Mao, Chengjian;Shapiro, David J.
雌激素通过雌激素受体α（ER α）起作用，刺激乳腺癌增殖，使ER α成为一个有吸引力的药物靶点。由于增殖抑制剂的384孔格式筛选对于某些细胞可能具有挑战性，因此基于内切酶的报告基因的抑制通常用作替代终点。为了鉴定17 β-雌二醇（E-2）-ER α刺激细胞增殖的新型小分子抑制剂，我们建立了一种基于细胞的雌激素反应元件（ERE）（3）-荧光素酶报告基因的E-2-ER α诱导抑制剂筛选。在分层随访测定中评价了75个“命中”，以鉴定命中未能进展的地方，并评价其作为E-2-ER α诱导的乳腺癌细胞增殖的抑制剂的有效性。来自荧光素酶筛选的75个命中中仅8个抑制ER α阳性MCF-7和T47 D细胞的雌激素诱导的增殖，但不抑制对照ER α阴性MDA-MB-231细胞。尽管12%的化合物仅在一种ER α阳性细胞系中抑制E-2-ER α刺激的增殖，但40%的化合物是有毒的并抑制所有细胞系的生长，并且类似于37%的化合物表现出很少或没有抑制E-2-ER α刺激的细胞增殖的能力。对代表性化合物进行了更详细的评价，并鉴定了一种主要的ER α抑制剂。
Estrogens, acting through estrogen receptor alpha (ER alpha), stimulate breast cancer proliferation, making ER alpha an attractive drug target. Since 384-well format screens for inhibitors of proliferation can be challenging for some cells, inhibition of luciferase-based reporters is often used as a surrogate end point. To identify novel small-molecule inhibitors of 17 beta-estradiol (E-2)-ER alpha-stimulated cell proliferation, we established a cell-based screen for inhibitors of E-2-ER alpha induction of an estrogen response element (ERE)(3)-luciferase reporter. Seventy-five "hits" were evaluated in tiered follow-up assays to identify where hits failed to progress and evaluate their effectiveness as inhibitors of E-2-ER alpha-induced proliferation of breast cancer cells. Only 8 of 75 hits from the luciferase screen inhibited estrogen-induced proliferation of ER alpha-positive MCF-7 and T47D cells but not control ER alpha-negative MDA-MB-231 cells. Although 12% of compounds inhibited E-2-ER alpha-stimulated proliferation in only one of the ER alpha-positive cell lines, 40% of compounds were toxic and inhibited growth of all the cell lines, and similar to 37% exhibited little or no ability to inhibit E-2-ER alpha-stimulated cell proliferation. Representative compounds were evaluated in more detail, and a lead ER alpha inhibitor was identified.