Discovery and restoration of aberrant nuclear structure and genome behaviour in breast cancer cells
Year: 2013
Type: dissertation
Abstract: The eukaryotic interphase nucleus is well organised and the genome positioned nonrandomly.Nuclear structure is an important regulator of genome behaviour and function.Genome organisation and nuclear structure are compromised in diseases such as cancer and laminopathies.This study was to find out and to determine if there is any functional relationship between nuclear structure and genome mis-organisation in cancer cells.I have assessed the presence and distribution of specific nuclear structural proteins (A-type, B-type lamins and its receptor LBR, many of their binding proteins such as MAN1, LAP2α, LAP2, and Emerin and other nuclear proteins (PML, Nucleolin, and Ki67) using indirect immunofluorescence.From this study, it is found that the nuclear structure of breast cancer cells is often altered.The most severely affected proteins are the nuclear lamins B1 and B2 and they found as large foci within the nucleoplasm with little LBR expression to localise the lamin B. I also assessed the chromosome positioning (HSA 7, 10, 11, 14 and 17) and gene positioning (AKT1, CCND1, HSP90AA1, EGFR, ERRBB2/HER2 and PTEN) in breast cancer cell lines (T-47D, GI-101, Sk-Br-3 and BT-474) and in normal breast cell lines (MCF-10A) using 2D-FISH technique.I also assessed the position of the genes in nuclei and correlated with gene expression using qRT-PCR.Breast cell lines have treated with a drug named lovastatin and it was found that the cells have restored LBR expression and localisation of lamin B, leading to altered gene positioning and changed expression of breast cancer genes.Since the drug (lovastatin, 12 µM/48 hours) affects the prenylation as a post-translation modification process and lamins B biosythensis, it is found that B-type lamins and its receptor expression and distribution were improved and increased in expression by 2-fold in expression levels in the most affected cells (T-47D, and BT-474) compared to the normal cells (MCF-10A) and these cells also showed abnormal nuclei and dead cells.When I would like to express my thanks to the Ministry of Higher Education and Scientific Research (Sudan), Gordon Memorial College Trust Fund (UK), Laboratory of Nuclear and Genomic Health (Biosciences
Author: Mai Hassan Ahmed
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: green
OpenAlex ID: W1523119066