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The effect of OTK18 upregulation in U937 cells on neuronal survival

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Abstract

The intent of this study was to characterize the effect OTK18 upregulation in monocytic cells had on neuronal survival. The human monocytic cell line, U937, was differentiated into macrophages or left as an undifferentiated monocyte. These cells were transfected with a plasmid containing the enhanced green fluorescent protein and OTK18 (pEGFP-OTK18) or an empty control vector (pEGFP-N3). The supernatants from the transfected U937 cells were used to culture rat neuronal cells (PC12). A live/dead assay was performed to determine the effect of culturing on cell survival. The protein levels of the neurotoxin, tumor necrosis factor alpha (TNF-α), and the neurotrophin, neurotrophin three (NT3), were determined by enzyme linked immunosorbent assay. The results of the live/dead assay showed differential cell survival between conditions with pEGFP-OTK18 when compared to the control empty vector. Quantitative real-time polymerase chain reaction assays demonstrated that OTK18 had an increased expression level when compared to the control. Lastly, NT3 protein levels were upregulated in treated cells with increased OTK18 expression, suggesting that OTK18 may play a role in neurotrophin production and consequently support neuronal survival.

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Acknowledgments

The authors thank Stephanie Larson for initial technical assistance. This research was supported by NIH grant no. 1 P20 RR16469 from the INBRE Program of the National Center for Research Resources, the UNK Research Services Council Undergraduate Student Research grant, UNK Office of Sponsored Programs (Summer Student Research Program), the UNK College of Natural and Social Sciences, the UNK Department of Graduate Studies and Research, and the UNK Biology Department.

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  1. Biology Department, University of Nebraska at Kearney, 905 W25th St., Kearney, NE, 68849, USA

    Christine E. Gilling & Kimberly A. Carlson

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  1. Christine E. Gilling
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  2. Kimberly A. Carlson
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Correspondence to Kimberly A. Carlson.

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Editor: J. Denry Sato

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Gilling, C.E., Carlson, K.A. The effect of OTK18 upregulation in U937 cells on neuronal survival. In Vitro Cell.Dev.Biol.-Animal 45, 243–251 (2009). https://doi.org/10.1007/s11626-009-9175-8

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  • DOI: https://doi.org/10.1007/s11626-009-9175-8

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