Browsing by Subject "proliferation"
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Item Effect of CD44 Expression on T Cell Acute Lymphocytic Leukemia(2014-08-01) Racine, Ronny R.; Mummert, Mark E.; Jones, Harlan; Berg, Rance E.CD44 is a cell surface glycoprotein that serves as a multifunctional receptor aiding in trafficking and adhesion of immune cells. CD44 also serves as a recruitment platform for signaling molecules and has been shown to regulate proliferation. In several types of leukemia the presence or absence of CD44 expression is associated with different clinical outcomes, with patients who have increased expression of CD44 exhibiting a stronger response to conventional chemo- and radiotherapy. By using Jurkat T cells, which do not express CD44, to determine the effects of CD44 expression in a model Acute Lymphocytic Leukemia cell line, we have outlined two major areas of study. Firstly, upon expression of CD44, Jurkat T cells proliferate slowly compared to the control cells. This decrease in proliferation is coupled to an arrest in the cell cycle during the transition from the G1 phase into S phase. The dysregulation of the cell cycle induced by CD44 also leads to the induction of aneuploidy. CD44 expressing Jurkat T cells have reduced mRNA expression for several key regulators of chromosome separation and the mitotic spindle complex. This finding, coupled with decreased EGR-1 expression, which controls the cyclins responsible for transition from G1 into S phase, leads to an unstable cell phenotype which proliferates slowly and accumulates extra chromosomes in daughter cells. The second area of study focuses on the mechanism by which CD44 expression at the cell surface results in the observed decreases in proliferation, Akt activation, and EGR-1 expression. We observed that CD44 expressing Jurkat cells show four to five times higher calcium influx when at rest compared to the vector control cells. This influx in calcium is due to CD44 expression activating a cell surface inducible calcium release activated calcium channel. The excess calcium activates calcium-activated phosphatases and kinases, disrupting EGR-1 expression and inducing a hypophosphorylation of Akt. Together, these findings indicate that CD44 expression can regulate cell proliferation and signal transduction pathways in addition to its role in adhesion. Thus, our data provide a further understanding of how CD44 expression modifies leukemic cells into cells that are favorable for therapeutic intervention.Item Endothelin-1-Induced Proliferation of Human Optic Nerve Head Astrocytes Under Hypoxia(2003-11-01) Desai, Devashish; Thomas Yorio; Ganesh Prasanna; Clark, Abbot F.Desai, Devashish, Endothelin-1-Induced Proliferation of Cultured Human Optic Nerve Head Astrocytes under Hypoxia. Master of Science (Biomedical Sciences). Purpose: Optic nerve head astrocytes (ONAs) normally support and protect the axons of retinal ganglion cells exiting the eye. Along with effects related to elevated intraocular pressure (IOP), proliferation and activation of ONAs, known as ‘astrogliosis’, is also thought to contribute to the pathophysiology of glaucoma by distributing axonal transport and preventing axon regeneration. Concentrations of endothelin-1 (ET-1) are elevated in glaucomatous eyes and in animal models for glaucoma. ET-1 injection into the eye causes reduction of ocular blood flow. ET-1 causes a time-dependent proliferation of human ONAs. Tumor necrosis factor-α (TNF-α), a cytokine, which is also elevated in glaucomatous optic nerve head, promotes ET-1 release from ocular cells and could potentially stimulate ET-1 secretion from the ONAs. Hypoxia resulting from ischemia, which is produced by the elevation of IOP or vasospasm in the retinal vasculature, is considered a significant factor contributing to the stress as the glaucomatous optic nerve head. Methods: Concentrations of ET-1 secreted by hONAs into cell culture media after hypoxia and TNF-α treatment was measured using an enzyme-linked immunosorbent assay (ELISA). Proliferation of hONAs was measured using a proliferation assay (formazan assay), performed at the end of various time periods of incubation with TNPα and ET-1 under normoxia or hypoxia. The involvement of mitogen activated protein kinase (MAPK) in hONA proliferation was examined using MAPK inhibitors and Western blot analyses. Results: Cell culture media collected from hONAs after 24-hour hypoxia with concurrent TNF-α treatment showed a 500% increase in the irET-1. Under normoxia, both TNF-α and ET-1 caused moderate proliferation of hONAs. Under hypoxia, TNF-α-induced proliferation was greatly increased. Conclusion: Hypoxia augments TNF-a and ET-1 growth of optic nerve head astrocytes, by way of increasing ET-1 synthesis and release as well as mitogenesis. Therefore reactive ONAs could be the common denominator underlying optic nerve damage in glaucoma since their localization makes them susceptible to mechanistic and ischemic influences in addition to influences of ET-1 and TNF-α. Keywords: astrocyte; endothelin-1; tumor-necrosis factor-α; hypoxia; proliferation; astrogliosis; glaucoma; optic nerve