胶质母细胞瘤（GBM）是最常见的中枢神经系统肿瘤，与不良预后相关。近几十年来，GBM死亡率没有显著改善。ER-α36是ER-α66的变体，可能通过基因组和非基因组机制参与癌瘤生长和增殖。该变异体可能在多种肿瘤的他莫昔芬（tamoxifen）耐药中扮演重要角色。我们实验室之前发现ER-α36在GBM中表达并参与增殖，但是ER-α36在GBM侵袭中的作用仍然未知。本研究旨在确定ER-α36调节剂SNG162对GBM生长和侵袭的影响。U251细胞、U87细胞和ER-α36表达减少的U87-36KD细胞在二维和三维（3D）环境中培养。通过细胞计数、流式细胞术、Western印迹和MTT测定法检查GBM细胞的增殖，并用共聚焦显微镜在3D环境中测定侵袭性。经处理1、3和5μM的SNG162后，表达EGFR和ER-α36减少的U87细胞生长显著减少；在U251细胞中，生长抑制作用比在U87细胞中更强，尽管U251细胞中ER-α36的表达水平低于U87细胞。我们发现1μM的SNG162抑制了U87细胞中E2-诱导的MAPK/ERK通路的激活。我们还展示了SNG162抑制了U87细胞的侵袭，但是它在使用3D培养方法时并没有显著影响U251和U87-36KD细胞的侵袭。最后，我们确定ER-α36在侵袭性GBM细胞的细胞核中表达，并且SNG162显著抑制了这些细胞中ER-α36的表达。SNG162抑制了非侵袭性GBM细胞膜上EGFR的表达。这些结果表明，SNG162可能是通过靶向ER-α36的治疗剂用于GBM的一个疗法。版权所有©2023 Elsevier Inc.

Glioblastoma (GBM) is the most common central nervous system tumor and is associated with poor outcomes. There have been no significant improvements in GBM mortality in recent decades. ER-α36 is a variant of ER-α66 that may be involved in carcinoma growth and proliferation via genomic and nongenomic mechanisms. This variant might play an essential role in tamoxifen resistance of several tumors. Previously, our laboratory found that ER-α36 is expressed in GBM and participates in proliferation; nevertheless, the role of ER-α36 in GBM invasion remains unknown. This study aimed to determine the effects of the ER-α36 modulator SNG162 on GBM growth and invasion. U251 cells, U87cells, and U87-36KD cells with knockdown of ER-α36 expression were cultured under the two-dimensional and the three-dimensional (3D) environments. GBM cells growth was examined by cell counting, flow cytometry, western blot, and MTT assays. Invasiveness was measured using confocal microscopy in the 3D environment. Growth of U87 cells with downregulated EGFR and ER-α36 expression was significantly reduced after treatment with 1 µM, 3 µM, and 5 µM of SNG162; growth inhibition in U251 cells was more potent than in U87 cells, although the expression level of ER-α36 in U251 cells was lower than in U87 cells. We found that 1 μM SNG162 suppressed E2-induced MAPK/ERK pathway activation in U87 cells. We also showed that SNG162 inhibited U87 cells invasion; however, it did not significantly affect U251 and U87-36KD cells invasion using the 3D culture method. Finally, we determined that ER-α36 was expressed in the nucleus of invading GBM cells, and SNG162 significantly inhibited the expression of ER-α36 in these cells. SNG162 inhibited the expression of EGFR on cell membranes of non-invasive GBM cells. These results suggest that SNG162 could be a therapeutic agent for GBM by targeting ER-α36.Copyright © 2023. Published by Elsevier Inc.