PDF(Pubmed)
Abstract：
Whether G protein-coupled estrogen receptor 1 (GPER1) is tumor-promoting or tumor-suppressive depends in part on tumor entity. Little is known about the function of GPER1 in vulvar carcinoma. In this work, we aim to clarify what role GPER1 plays in vulvar cancer, tumor-promoting or tumor-suppressive. Localization of GPER1 in A431 and CAL-39 vulvar carcinoma cells was examined by immunofluorescence. Using a tissue microarray of vulvar neoplasias, the correlation between GPER1 expression and grade of malignancy was investigated. A431 and CAL-39 cells were treated either with GPER1 agonist G1 or antagonist G36. Proliferation was quantified by BrdU assay and viability examined using Resazurin assay. Morphological changes were analyzed by microscopy and measured using ImageJ. Cell migration was analyzed by gap closure assay. Clonogenic potential was tested by colony and sphere formation. Expression of estrogen receptors was examined by Western blot. GPER1 was found consistently expressed in vulvar neoplasia tissues. The immune-reactive score was found to be significantly higher in tissue samples of lymph node metastases and neoplasias with grade 3. In A431 and CAL-39 vulvar carcinoma cells, GPER1 expression was mainly found in the cytoplasm and nuclei. Treatment of A431 and CAL-39 cells with GPER1 agonist G1 resulted in a decrease in proliferation and migration. In addition, colony formation and tumor sphere formation were reduced. Furthermore, morphological signs of necrosis and reduction in cell viability after G1 treatment were observed. The GPER1 antagonist G36 did not have significant effects on vulvar carcinoma cells. Neither agonist G1 nor antagonist G36 treatment resulted in altered expression of estrogen receptors. Activation of GPER1 with GPER1 agonist G1 reduces the tumorigenic potential of the vulvar carcinoma cells. It can be deduced from this that GPER1 appears to have a tumor-suppressive effect in vulvar carcinoma.
摘要：
G蛋白偶联雌激素受体1（GPER1）是促进肿瘤还是抑制肿瘤部分取决于肿瘤实体。关于GPER1在外阴癌中的功能知之甚少。在这项工作中,我们的目的是阐明GPER1在外阴癌中的作用，促进肿瘤或抑制肿瘤。通过免疫荧光检查GPER1在A431和CAL-39外阴癌细胞中的定位。使用外阴肿瘤的组织微阵列，研究了GPER1表达与恶性肿瘤分级之间的相关性.用GPER1激动剂G1或拮抗剂G36处理A431和CAL-39细胞。通过BrdU测定定量增殖,并使用Resazurin测定检查活力。通过显微镜分析形态学变化并使用ImageJ测量。通过间隙闭合测定分析细胞迁移。通过菌落和球体形成来测试克隆生成潜力。通过Western印迹检查雌激素受体的表达。发现GPER1在外阴瘤形成组织中一致表达。在淋巴结转移和3级肿瘤的组织样本中发现免疫反应评分明显更高。在A431和CAL-39外阴癌细胞中，GPER1表达主要见于细胞质和细胞核。用GPER1激动剂G1处理A431和CAL-39细胞导致增殖和迁移减少。此外,集落形成和肿瘤球形成减少。此外，观察到G1处理后坏死和细胞活力降低的形态学迹象。GPER1拮抗剂G36对外阴癌细胞无明显影响。激动剂G1和拮抗剂G36治疗均未导致雌激素受体表达改变。用GPER1激动剂G1激活GPER1可降低外阴癌细胞的致瘤潜能。由此可以推断，GPER1似乎在外阴癌中具有肿瘤抑制作用。
