Abstract

Cell membrane is the biological barrier serving as both territorial defense and the communication hinge for the interior of cell from its surroundings. As building blocks of cellular membranes and also precursor for second messengers, a variety of lipids play essential roles in cellular membrane dynamics as well as important functions such as cell proliferation, apoptosis, signal transduction and membrane trafficking modulation. Lipidomics, representing the systematic and integrative studies of diversified lipids (lipidome) in a biological system, is an emerging yet rapid developing field and hence requires advanced and complementary analytical techniques as well as multiple statistical tools. Our development of reliable analytical methodology (the advanced Mass Spectrometric and high-resolution NMR techniques) and application of multiple statitistical approaches (multivariate data analysis and univariate t-test) enable us to achieve these comprehensive understandings. We have investigated, first of all, the effects induced by hypoxia on cervical cancer derived cells (HeLa cells) to see how and how much the changes in phospholipids profile are able to get light into the targeted biological problem (hypoxia) and provide a preliminary insight into the underlying mechanisms. We found that hypoxia stimulation dramatically reduced the total amount of cellular phosphoinositols (PI) but prominently increased the amount of lyso phosphocholines (lyso-PC) and lyso phosphoethanolamines (lyso-PE). Moreover, our studies suggested the polyunsaturated phospholipids species as stronger biomarkers upon hypoxia treatment. The evaluation of changes in the average unsaturation index (UI) of the membrane lipids acyl chains revealed that UI slightly increased in several lipid classes, thus affecting membrane fluidity and further membrane-dependent functions. The plausible mechanisms by HeLa cells to adapt to hypoxia conditions are briefly reported as well. We have also conducted the comparative lipidomic studies of urothelial cancer cell line RT4 (a model system of a benign tumor) and T24 (a model system of a metastatic tumor) aiming to reveal probable roles and relevant differential changes of membrane lipids with respect to urinary bladder metastasis progress. Significant changes of lipids metabolism were found to correlate with urothelial nonmetastatic and metastatic cell models. The most remarkable finding was that the malignant cell type (T24) showed a strong decrease of ether PC species complemented by a sharp increase of the length and the average unsaturation number of lipids acyl chains. Ceramide-based sphinglipids also showed altered profiles in these two cell types. Such analyses suggest a certain significant re-organization of cellular membrane in malignant cell transformation, involving variations in compositional lipid structures and possible signaling transduction pathways. Observations of such reduction of the 1-alkyl PC species and the chain shortening of lipid species might serve as a tool in urinary bladder cancer intervention.