The In Vitro Effect of Ethanol on Rat Lingual Epithelium

Bell, Aileen (1993) The In Vitro Effect of Ethanol on Rat Lingual Epithelium. PhD thesis, University of Glasgow.

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Chronic alcohol abuse has been linked to the development of oral cancer, but before studying the role of alcohol in intra-oral carcinogenesis the effect of alcohol alone upon the oral epithelium must be clarified. Since cells express their individuality through the proteins they produce, alcohol related alterations are likely to be mediated or accompanied by alterations in the epithelial protein pattern. Graham & Rennie (1987) noted alterations in the levels of three proteins from rat lingual epithelium after 102 days of alcohol intake. However, animal models are time consuming and expensive. It was thus the aim of the work reported in this thesis to develop an in vitro model to investigate the effects of alcohol on rat lingual epithelium and to compare any changes in epithelial protein profile with those described in vivo. The results of this study showed that levels of the 66-70kD protein increased at 12 and 24 hours after heat treatment. Heat treatment did not alter the levels of the other four proteins altered by alcohol and acetaldehyde. The increase in levels of the 66-70kD protein appears to be a general stress response while the alterations in levels of the other proteins are specifically related to alcohol/acetaldehyde. The possibility that the 66-70kD protein was a HSP was further investigated using immunofluorescent staining with an anti-HSP 70kD antibody. Acetaldehyde treated cells and cells at 12 and 24 hours after heat treatment stained positively. A Western blot carried out 24 hours after heat treatment using the anti-HSP 70kD antibody detected a positive band in the 70kD region. Simple subcellular localisation studies using differential centrifugation and Triton X-100 detergent lysis of cells showed that the three higher MW proteins were present in the insoluble cell fraction while the two lower MW proteins were present in the soluble cell fraction. The significance of the increased levels of the 30kD and 28kD proteins is not clear, although they do not appear to be HSPs. The 66-70kD protein appears to be a HSP and would therefore assist the epithelial cells resist external stress. The high molecular weights and the presence of the 230kD and 200kD proteins in the insoluble cell fraction suggests that they may be large structural proteins, and as such are probably keratin peptides and polypeptides. The work presented in this thesis confirms earlier in vivo studies (Graham, 1987; Graham & Rennie, 1987) and indicates that the in vitro model is a useful system for the future study of the effects of alcohol on oral epithelium. This work has also described the importance of acetaldehyde and emphasises the need for further investigation of the role of acetaldehyde in alcohol related disease processes. (Abstract shortened by ProQuest.).

Item Type: Thesis (PhD)
Qualification Level: Doctoral
Additional Information: Adviser: J S Rennie
Keywords: Medicine, Toxicology
Date of Award: 1993
Depositing User: Enlighten Team
Unique ID: glathesis:1993-74709
Copyright: Copyright of this thesis is held by the author.
Date Deposited: 13 Nov 2019 15:58
Last Modified: 13 Nov 2019 15:58

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