PDF(Pubmed)
Abstract：
Aquaporin (AQP) channels in endometrial cancer (EC) cells are of interest as pharmacological targets to reduce tumor progression. A panel of compounds, including AQP1 ion channel inhibitors (AqB011 and 5-(phenoxymethyl) furan-2-carbaldehyde, PMFC), were used to test the hypothesis that inhibition of key AQPs can limit the invasiveness of low- and high-grade EC cells. We evaluated the effects on transwell migration in EC cell lines (Ishikawa, MFE-280) and primary EC cells established from surgical tissues (n = 8). Quantitative PCR uncovered classes of AQPs not previously reported in EC that are differentially regulated by hormonal signaling. With estradiol, Ishikawa showed increased AQPs 5, 11, 12, and decreased AQPs 0 and 4; MFE-280 showed increased AQPs 0, 1, 3, 4, 8, and decreased AQP11. Protein expression was confirmed by Western blot and immunocytochemistry. AQPs 1, 4, and 11 were colocalized with plasma membrane marker; AQP8 was intracellular in Ishikawa and not detectable in MFE-280. AQP1 ion channel inhibitors (AqB011; PMFC) reduced invasiveness of EC cell lines in transwell chamber and spheroid dispersal assays. In Ishikawa cells, transwell invasiveness was reduced ~41% by 80 µM AqB011 and ~55% by 0.5 mM 5-PMFC. In MFE-280, 5-PMFC inhibited invasion by ~77%. In contrast, proposed inhibitors of AQP water pores (acetazolamide, ginsenoside, KeenMind, TGN-020, IMD-0354) were not effective. Treatments of cultured primary EC cells with AqB011 or PMFC significantly reduced the invasiveness of both low- and high-grade primary EC cells in transwell chambers. We confirmed the tumors expressed moderate to high levels of AQP1 detected by immunohistochemistry, whereas expression levels of AQP4, AQP8, and AQP11 were substantially lower. The anti-invasive potency of AqB011 treatment for EC tumor tissues showed a positive linear correlation with AQP1 expression levels. In summary, AQP1 ion channels are important for motility in both low- and high-grade EC subtypes. Inhibition of AQP1 is a promising strategy to inhibit EC invasiveness and improve patient outcomes.
摘要：
子宫内膜癌(EC)细胞中的水通道蛋白(AQP)通道作为减少肿瘤进展的药理学靶标是令人感兴趣的。一组化合物，包括AQP1离子通道抑制剂（AqB011和5-（苯氧基甲基）呋喃-2-甲醛，PMFC),用于检验以下假设：关键AQPs的抑制可以限制低级和高级EC细胞的侵袭性。我们评估了对EC细胞系中transwell迁移的影响（Ishikawa，MFE-280）和从手术组织建立的原代EC细胞（n=8）。定量PCR揭示了以前在EC中未报道的受激素信号传导差异调节的AQPs类别。有了雌二醇,Ishikawa显示AQPs5、11、12增加，AQPs0和4减少；MFE-280显示AQPs0、1、3、4、8增加，AQP11减少。蛋白质表达通过蛋白质印迹和免疫细胞化学证实。AQPs1、4和11与质膜标记物共同定位；AQP8在Ishikawa细胞内，在MFE-280中检测不到。AQP1离子通道抑制剂（AqB011;PMFC）在transwell室和球体分散测定中降低了EC细胞系的侵袭力。在石川牢房里，transwell的侵袭力降低了约41%，降低了80µMAqB011，降低了约55%，降低了0.5mM5-PMFC。在MFE-280中,5-PMFC抑制侵袭约77%。相比之下,拟议的AQP水孔抑制剂(乙酰唑胺，人参皂苷,KeenMind,TGN-020，IMD-0354）无效。用AqB011或PMFC处理培养的原代EC细胞显着降低了transwell腔室中低级和高级原代EC细胞的侵袭力。我们证实了通过免疫组织化学检测的肿瘤表达中度至高水平的AQP1，而AQP4、AQP8和AQP11的表达水平显著降低。AqB011治疗EC肿瘤组织的抗侵袭能力与AQP1表达水平呈正线性相关。总之,AQP1离子通道对于低级和高级EC亚型的运动性都很重要。抑制AQP1是抑制EC侵袭性和改善患者预后的有希望的策略。
