Inhibition of the phosphatidylinositol-3-kinase pathway abrogates polyinosinic:polycytidylic acid-stimulated hyaluronan-mediated human mucosal smooth muscle cell binding of U937 monocytic cells.
The origin of inflammatory bowel disease (IBD) is unknown and likely to be multifactorial. Our laboratory has established that in human mucosal smooth muscle cells (M-SMCs), cellular stress induced by virus or the viral mimic double-stranded RNA (polyinosinic:polycytidylic acid [poly I:C]) increases cell surface hyaluronan (HA) deposition and the formation of long cable-like structures of HA that are important for leukocyte attachment. Since leukocyte accumulation and hyperplasia of the M-SMCs are characteristic pathological changes observed in IBD patients, and phosphatidylinositol-3-kinase (PI3K)/Akt signaling pathways play established roles in cell survival, we investigated whether this pathway is involved in this unique HA-mediated leukocyte attachment. Poly I:C-stimulated M-SMCs bind significantly more monocytic cells than untreated cells and this response was inhibited in a dose-dependent manner by treatment with the PI3K inhibitor, LY294002. Since Akt is a critical downstream regulator of PI3K, we investigated the phosphorylation status of Akt in M-SMCs after treatment with poly I:C for 1 h and found that Akt was phosphorylated, but the phosphorylated Akt band was undetectable in LY294002 plus poly I:C-treated cultures. Confocal microscopy of M-SMCs stained for HA revealed that HA cable formation after poly I:C treatment was abrogated by LY294002. These results demonstrate that poly I:C-stimulated M-SMCs phosphorylate Akt, produce HA cables, and promote HA-mediated leukocyte adhesion through a PI3K/Akt-dependent manner.
Bandyopadhyay
Sudip K
SK
Department of Pathobiology, Lerner Research Institute , Cleveland Clinic, Cleveland, Ohio 44195, USA. bandyos@ccf.org
de la Motte
Carol A
CA
Majors
Alana K
AK
Strong
Scott A
SA
D002448:Cell Adhesion / Q000187:drug effects / Q000276:immunology; D002867:Chromones / Q000494:pharmacology; D006801:Humans; D006820:Hyaluronic Acid / Q000276:immunology / Q000378:metabolism*; D018928:Immunity, Mucosal; D015212:Inflammatory Bowel Diseases / Q000188:drug therapy / Q000276:immunology* / Q000378:metabolism; D009000:Monocytes / Q000187:drug effects / Q000276:immunology / Q000378:metabolism* / Q000473:pathology; D009025:Morpholines / Q000494:pharmacology; D032389:Myocytes, Smooth Muscle / Q000187:drug effects / Q000276:immunology / Q000378:metabolism* / Q000473:pathology; D000081082:Phosphoinositide-3 Kinase Inhibitors; D011070:Poly I-C / Q000276:immunology / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects / Q000276:immunology; D020298:U937 Cells
D002867:Chromones; D009025:Morpholines; D000081082:Phosphoinositide-3 Kinase Inhibitors; C085911:2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one; D006820:Hyaluronic Acid; D011070:Poly I-C