NMDA receptors are expressed in human ovarian cancer tissues and human ovarian cancer cell lines.
NMDA receptors are expressed in human ovarian cancer tissues and human ovarian cancer cell lines.
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DOI：
10.2147/cpaa.s90367
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发表时间：
2015
期刊：
影响因子：
--
通讯作者：
Akerman B
中科院分区：
文献类型：
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作者：
North WG;Liu F;Tian R;Abbasi H;Akerman B
我们先前已经证明，乳腺癌和小细胞肺癌表达功能性NMDA受体，可以靶向促进癌细胞死亡。人卵巢癌组织和人卵巢癌细胞系（SKOV 3、A2008和A2780）也表达NMDA受体亚单位1（GluN 1）和亚单位2B（GluN 2B）。十七卵巢癌在两个阵列中进行了筛选免疫组化使用多克隆抗体，识别细胞外部分的GluN 1和GluN 2B。这些标本包括病理诊断为浆液性乳头状囊腺癌、类囊腺癌和透明细胞癌的恶性组织。此外，还评价了在该研究所治疗的10名患者的定义为卵巢腺癌的存档组织。所有的癌组织表现出阳性染色模式与NMDA受体抗体，而没有染色的肿瘤附近的正常组织或正常卵巢组织切片。Western blotting结果显示人卵巢腺癌细胞系（A2008、A2780、SKOV 3）均表达GluN 1，但表达水平不同。通过利用GluN 1抗体的免疫细胞化学和使用共聚焦显微镜成像，我们能够证明GluN 1蛋白在这些细胞的表面上表达。除了这些发现之外，使用针对GluN 2B蛋白的多克隆抗体证明了该蛋白的表达。发现用抗GluN 1抗体处理所有卵巢细胞系会导致细胞活力下降（P<0.001），下降至未处理细胞的10%-25%。用各种稀释度的GluN 1抗体处理对照HEK 293细胞对细胞活力没有影响。GluN 1拮抗剂MK-801（马来酸地佐环平）和GluN 2B拮抗剂ifenprodil可显著降低卵巢癌A2780细胞的存活率（P<0.01）。用艾芬地尔（2.5mg/kg体重/天）处理A2780肿瘤异种移植物显著降低nu/nu小鼠中的肿瘤生长。我们的研究结果表明，GluN 1和GluN 2B蛋白作为膜组分可以很容易地用于治疗大多数卵巢癌的目标。
We have earlier demonstrated that breast cancer and small-cell lung cancer express functional NMDA receptors that can be targeted to promote cancer cell death. Human ovarian cancer tissues and human ovarian cancer cell lines (SKOV3, A2008, and A2780) have now been shown to also express NMDA-receptor subunit 1 (GluN1) and subunit 2B (GluN2B). Seventeen ovarian cancers in two arrays were screened by immunohistochemistry using polyclonal antibodies that recognize an extracellular moiety on GluN1 and on GluN2B. These specimens comprised malignant tissue with pathology diagnoses of serous papillary cystadenocarcinoma, endometrioid adenocarcinoma, and clear-cell carcinoma. Additionally, archival tissues defined as ovarian adenocarcinoma from ten patients treated at this institute were also evaluated. All of the cancerous tissues demonstrated positive staining patterns with the NMDA-receptor antibodies, while no staining was found for tumor-adjacent normal tissues or sections of normal ovarian tissue. Human ovarian adenocarcinoma cell lines (A2008, A2780, SKOV3) were demonstrated to express GluN1 by Western blotting, but displayed different levels of expression. Through immunocytochemistry utilizing GluN1 antibodies and imaging using a confocal microscope, we were able to demonstrate that GluN1 protein is expressed on the surface of these cells. In addition to these findings, GluN2B protein was demonstrated to be expressed using polyclonal antibodies against this protein. Treatment of all ovarian cell lines with antibodies against GluN1 was found to result in decreased cell viability (P<0.001), with decreases to 10%–25% that of untreated cells. Treatment of control HEK293 cells with various dilutions of GluN1 antibodies had no effect on cell viability. The GluN1 antagonist MK-801 (dizocilpine maleate) and the GluN2B antagonist ifenprodil, like antibodies, dramatically decreased the viability of A2780 ovarian tumor cells (P<0.01). Treatment of A2780 tumor xenografts with ifenprodil (2.5 mg/kg body weight/day) significantly reduced tumor growth in nu/nu mice. Our findings suggest that both GluN1 and GluN2B proteins as membrane components could be readily available targets for the treatment of most ovarian cancers.