Chemical modification of cell proliferation and fluid secretion in renal cysts.
We used an in vitro model, MDCK cyst, to determine the extent to which pharmacologic compounds known to inhibit plasma membrane solute transport mechanisms could alter the enlargement of renal epithelial cysts. Solitary MDCK cells cultured within collagen gel undergo clonal growth to form true epithelial cysts in which a single layer of polarized cells (apex toward lumen) encloses a fluid-filled cavity. Repeated observations by light microscopy were used to quantitate the rate of cyst growth in diameter, and demonstrated that cyst enlargement involved an increase in cell number (proliferation) and a net increase in intracystic volume (fluid secretion). Intracyst pressure was greater than the interstitium (6.7 mm H2O +/- 3.1 SD), indicating that fluid entry was secondary to net solute accumulation. Amiloride and seven amiloride analogs that inhibited to different degrees conductive Na+ transport, Na+-dependent H+ transport and Na+-dependent Ca++ transport reversibly decreased the rate of cyst enlargement. The effectiveness of these agents to retard cyst enlargement correlated with their relative potencies to inhibit Na+-dependent Ca++ transport. Morphologic examination indicated that amiloride and amiloride analogs decreased cell proliferation and fluid secretion to the same degree. Ouabain and vanadate (Na+K,ATPase inhibitors), and L-645,695 (Na+-dependent Cl-/HCO3- inhibitor) potently slowed cyst expansion. In contrast to amiloride and amiloride analogs, these agents caused an unusual degree of cellular stratification within the cyst walls, a finding consistent with the notion that fluid secretion was inhibited to a greater extent then cellular proliferation. We conclude that chemical inhibitors of primary and secondary active solute transport can diminish or halt the enlargement of epithelial cysts in vitro by decreasing the rate of cellular proliferation and/or net fluid secretion.
Grantham
J J
JJ
Department of Medicine, University of Kansas School of Medicine, Kansas City.
Uchic
M
M
Cragoe
E J
EJ
Kornhaus
J
J
Grantham
J A
JA
Donoso
V
V
Mangoo-Karim
R
R
Evan
A
A
McAteer
J
J
D000584:Amiloride / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D003470:Culture Media; D004847:Epithelial Cells; D004848:Epithelium / Q000187:drug effects / Q000648:ultrastructure; D006133:Growth Substances / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D008854:Microscopy, Electron; D008954:Models, Biological; D009994:Osmolar Concentration; D010042:Ouabain / Q000494:pharmacology; D007690:Polycystic Kidney Diseases / Q000473:pathology*
D003470:Culture Media; D006133:Growth Substances; D010042:Ouabain; D000584:Amiloride