Wilson P D, Hreniuk D, Gabow P A
Department of Physiology and Biophysics, UMDNJ-Robert Wood Johnson, Medical School, Piscataway 08854-5635.
J Cell Physiol. 1992 Feb;150(2):360-9. doi: 10.1002/jcp.1041500220.
Human autosomal dominant polycystic kidney disease (ADPKD) epithelia were grown in primary monolayer cultures and their properties compared with intact kidney epithelial cultures derived from individually microdissected normal human kidney proximal convoluted tubules (PCT), proximal straight tubules (PST), and cortical collecting tubules (CCT). In vivo, ADPKD cyst epithelia exhibited a thickened basement membrane, and immunofluorescence demonstrated the presence of laminin, fibronectin, type IV collagen, and heparan sulfate proteoglycan in basement membranes and type I collagen in the interstitium. ADPKD epithelia grown in culture synthesized and secreted basally a unique, extracellular matrix that took the form of proteinaceous spheroids when the cells were grown on dried, type I collagen. Incorporation of H2[S35O4] into basement membrane extracts was increased more than ten-fold in ADPKD epithelia by comparison to normal PST and CCT. In addition to incorporation into the normal tubular basement membrane 220 kD band, radioactivity was also seen at 175 kD and 150 kD in ADPKD extracts. Growth in culture of cyst-lining ADPKD epithelia was more rapid than normal tubules, and was abnormal since there was no absolute requirement for added extracellular matrix. However, when ADPKD epithelia were grown on different, exogenous matrix protein components, a profound influence on both structure and epithelial cell proliferation was seen. Growth on a complete basement membrane three-dimensional gel derived from the Engelbreth-Holm-Swarm (EHS) sarcoma led to a reduction in the numbers of spheroids and increase in amorphous filaments. Incorporation of [3H]-thymidine into ADPKD epithelia was greater than into normal PCT, PST, and CCT and was also greatly modified by the type of extracellular matrix components provided. In studies using single matrix components, the strongest proliferative response was seen when ADPKD epithelia were plated on type I collagen greater than type IV collagen greater than fibronectin greater than laminin. These findings suggest that the excessive growth of cyst-lining epithelia may be, at least in part, a result of abnormal basement membrane and extracellular matrix production by ADPKD cells.
人常染色体显性多囊肾病（ADPKD）上皮细胞在原代单层培养物中生长，并将其特性与源自个体显微切割的正常人肾近端曲管（PCT）、近端直管（PST）和皮质集合管（CCT）的完整肾上皮细胞培养物进行比较。在体内，ADPKD囊肿上皮细胞表现出基底膜增厚，免疫荧光显示基底膜中有层粘连蛋白、纤连蛋白、IV型胶原和硫酸乙酰肝素蛋白聚糖，间质中有I型胶原。在培养中生长的ADPKD上皮细胞在基底分泌一种独特的细胞外基质，当细胞在干燥的I型胶原上生长时，这种基质呈蛋白质球体形式。与正常PST和CCT相比，ADPKD上皮细胞中H2[S35O4]掺入基底膜提取物中的量增加了十多倍。除了掺入正常肾小管基底膜的220kD条带外，在ADPKD提取物中还在175kD和150kD处检测到放射性。囊肿内衬ADPKD上皮细胞在培养中的生长比正常肾小管更快，并且是异常的，因为对添加的细胞外基质没有绝对需求。然而，当ADPKD上皮细胞在不同的外源性基质蛋白成分上生长时，对结构和上皮细胞增殖都有深远影响。在源自恩格尔布雷特-霍尔姆-斯旺（EHS）肉瘤的完整基底膜三维凝胶上生长导致球体数量减少和无定形细丝增加。[3H]-胸苷掺入ADPKD上皮细胞中的量大于掺入正常PCT、PST和CCT中的量，并且也受到所提供的细胞外基质成分类型的极大影响。在使用单一基质成分的研究中，当ADPKD上皮细胞接种在I型胶原上时，观察到最强的增殖反应，其次是IV型胶原、纤连蛋白、层粘连蛋白。这些发现表明，囊肿内衬上皮细胞的过度生长可能至少部分是ADPKD细胞异常产生基底膜和细胞外基质的结果。
