Snijders, Kirsten (2016) In vivo and in vitro applications of biospectroscopy:potential use as a diagnostic tool which can provide insights into cell cycle dependent proliferation. Masters thesis, UNSPECIFIED.
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Abstract
Biospectroscopy and its applications characterise in vitro and in vivo cells with abnormal proliferation capacities. Proliferation of endometrial tissues in regions other than the uterus lining is known as endometriosis. It affects around 10% of women in the UK and worldwide. This study tests Fourier-Transform Infrared Attenuated Total Reflectance (FTIR-ATR) and Raman Spectroscopy as a novel approach for early identification and in depth classification of Endometriosis. An implementation of a non-invasive and high-throughput screening method would eliminate bias and thus reduce screening costs. The potential for double screening endometriosis and cervical cancer simultaneously offers a cost effective approach. Each patient spectra can be added to the data base to create a thorough in-depth identification method for varying disease morphologies. Results showed that both spectral techniques successfully detected endometriosis presence when the right pre-processing methods were applied, although Raman spectroscopy carried a higher degree of sensitivity and was able to detect ovarian endometriosis alongside endometriosis. Thus far this study demonstrates that Benzoapyrene (B[a]P) is capable of inducing alterations in exposed Michigan Cancer Foundation-7 (MCF-7) cell populations even at very low doses of 10-9M. B[a]P gradually reduces cell survival after 7 days exposure and induces long-term inhibition of cell growth. High dose exposure of 10-6M B[a]P causes short-term evasion of G1 arrest and accumulation in S phase. These findings indicate that growth kinetics in vitro are important pre-determinants of MCF-7 cell susceptibility to genotoxic agents. Biospectroscopy and computational analysis produced biochemical signatures characterising dose and cell cycle related effects of B[a]P that indicated DNA alterations being induced. Overall the cell cycle is an important factor in genotoxic synthesis and cell proliferation. These findings can be linked to abnormal tumour growth in cancer.