Determining the cellular localisation of novel cancer therapeutics

Rowley, Laura Elizabeth (2015). Determining the cellular localisation of novel cancer therapeutics. University of Birmingham. Ph.D.

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Abstract

Three areas of investigation regarding a novel class of anti-cancer therapeutics, triple-helicate dinuclear compounds known as ‘cylinders’, have been addressed.

An improved protocol for the synthesis of a ruthenium cylinder has been developed, utilising microwave synthesis rather than reflux techniques. The use of Sephadex C-25 as a solid phase for column chromatography led to shorter purification protocols.

Experiments to determine the cellular localisation of fluorescent ruthenium cylinder using confocal microscopy showed fluorescence within MDA-MB-231 cells, but not within SKOV-3 cells, suggesting preferential uptake between cell lines. Co-localisation experiments suggested localisation of the cylinder within cell nuclei.

Synchrotron radiation has been used to image iron cylinder within SKOV-3 cells. High levels of iron are found near the cell membrane, corresponding to areas of high calcium concentration. XANES spectra show that the iron is in an environment closer to that of ferric iron as opposed to the ferrous iron found within the cylinder.

The cellular effects of cylinder treatment have been imaged, showing that cell motility is compromised by cylinder treatment. Iron cylinder causes production of NO within MDA-MB-231 cells. Control images showed low levels of fluorescence within MDA-MB-231 cells, suggesting the presence of endogenous NO, which had been debated within literature.

Type of Work:

Thesis (Doctorates > Ph.D.)

Award Type:

Doctorates > Ph.D.

Supervisor(s):