Effects of tumour necrosis factor on BrdU incorporation in cultured human enterocytes.
Citation:J. McDevitt, C. Feighery, C. O?Farrelly, G. Martin, D.G. Weir, D.K. Kelleher `Effects of tumour necrosis factor on BrdU incorporation in cultured human enterocytes? in The Journal of Biological Chemistry, 4, 1995, 31-37
Effects of tumour necrosis factor-alpha on BrdU incorporation in cultured human enterocytes.pdf (published (publisher copy) peer-reviewed) 868.0Kb
Bromodeoxyuridine incorporation is a useful method for studying the pattern of DNA synthesis in proliferating cells. The distribution pattern of incorporated BrdU in villus enterocytes of duodenal explants was analysed after exposure to TNFalpha in organ culture. TNFalpha caused a consistent, low level uptake of BrdU in the portion of the nucleus close to the nuclear membrane, this pattern was absent from the control cultures. As these epithelial cells are terminally arrested in G(0), the BrdU incorporation was thought not to be due to S phase DNA synthesis, but rather a response to the cytotoxic influence of TNFalpha. Microtitre plate proliferation assays of cell density and DNA synthesis were devised to study the effects of TNFalpha on confluent monolayers of the human foetal jejunal cell line I407 and the mouse fibrosarcoma cell line L929. Both cell lines showed a similar response to TNFalpha. Exposure to TNFalpha alone did not reduce cell numbers but did cause a significant increase in DNA synthesis (p < 0.05). When cycloheximtde was added in tandem with TNFalpha there was a significant reduction in cell number (p < 0.001) and level of DNA synthesis (p < 0.01) indicative of cell death. The DNA of cells exposed to TNFalpha and cycloheximide was fragmented when viewed on an electrophoresis gel. The results show that BrdU incorporation might be a good indicator of damage to the DNA of cells after cytotoxic insult. TNFalpha may be responsible for villus enterocyte damage in enteropathies such as coeliac disease and GVHR of the small bowel.
Health Research Board
Publisher:Taylor and Francis
Type of material:Journal Article
Series/Report no:The Journal of Biological Chemistry
Availability:Full text available