Kidney Genetics Group, Academic Nephrology Unit, The Henry Wellcome Laboratories for Medical Research, University of Sheffield Medical School, Sheffield, United Kingdom.
Am J Physiol Renal Physiol. 2011 Jun;300(6):F1375-84. doi: 10.1152/ajprenal.00482.2010. Epub 2011 Mar 23.
Thiazolidinediones have been reported to retard cystic disease in rodent models by uncertain mechanisms. We hypothesized that their major effect in retarding cystogenesis was through inhibiting cell proliferation or stimulating apoptosis. In the Madin-Darby canine kidney cell (MDCK) model, rosiglitazone inhibited cyst growth in a time- and dose-dependent manner and this was accompanied by a reduction in basal proliferation and an increase in apoptosis. Unexpectedly, we also observed a striking abnormality in lumen formation resulting in a characteristic multiple lumen or loss of lumen phenotype in treated cells at doses which did not inhibit cell proliferation. These changes were preceded by mislocalization of gp135 and Cdc42, misorientation of the mitotic spindle, and retardation in centrosome reorientation with later changes in primary cilia length and mislocalization of E-cadherin. Cdc42 activation was unaffected by rosiglitazone in monolayer culture but was profoundly inhibited in three-dimensional culture. MDCK cells stably expressing mutant Cdc42 showed a similar mislocalization of gp135 expression and multilumen phenotype in the absence of rosiglitazone. We conclude that rosiglitazone influences MDCK cyst growth by multiple mechanisms involving dosage-dependent effects on proliferation, spindle orientation, centrosome migration, and lumen formation. Correct spatial Cdc42 activation is critical for lumen formation, but the effect of rosiglitazone is likely to involve both Cdc42 and non-Cdc42 pathways.
噻唑烷二酮类药物已被报道通过不确定的机制延缓啮齿动物模型中的囊性疾病。我们假设它们延缓囊发生的主要作用是通过抑制细胞增殖或刺激细胞凋亡。在马迪恩-达比犬肾细胞(MDCK)模型中,罗格列酮以时间和剂量依赖的方式抑制囊肿生长,这伴随着基础增殖减少和凋亡增加。出乎意料的是,我们还观察到管腔形成中的明显异常,导致在不抑制细胞增殖的剂量下处理细胞出现特征性的多个管腔或管腔缺失表型。这些变化之前伴随着 gp135 和 Cdc42 的定位错误、有丝分裂纺锤体的定向错误以及中心体重定向的延迟,随后初级纤毛长度和 E-钙粘蛋白的定位错误发生变化。Cdc42 激活在单层培养中不受罗格列酮影响,但在三维培养中受到强烈抑制。稳定表达突变 Cdc42 的 MDCK 细胞在没有罗格列酮的情况下也表现出 gp135 表达和多腔表型的类似定位错误。我们得出结论,罗格列酮通过多种机制影响 MDCK 囊肿生长,包括对增殖、纺锤体定向、中心体迁移和管腔形成的剂量依赖性影响。正确的空间 Cdc42 激活对于管腔形成至关重要,但罗格列酮的作用可能涉及 Cdc42 和非 Cdc42 途径。