| The function of the cell membrane protein will affected by the fluidity of lipid,however cholesterol is an important composition of micro-domains and it will affect the fluidity of cell membrane,so as to affect the function of proteins.Lipid rafts are cholesterol-and sphingophospholipids-enriched micro-domains on plasma membrane surface of mammal cells.The micro-domains are involved in a variety of cellular functions,including the regulation of cell adhesion and membrane signaling through proteins within lipid rafts,which lead to the development of novel targets for cancer therapy.Therefore,it has important significance to study the effects of cholesterol on plasma membrane lipid order in cancer cells.Fluorescence Correlation Spectroscopy(FCS)was employed to study the effects of cholesterol on the dynamics of plasma membrane lipid in MCF-7 cells quantitatively,in order to further study the processes of micro-domain participated in protein function.First of all,we stated the basic principle of fluorescence correlation spectroscopy,and tested the effectiveness of the systems by solution experiment.Then we used FCS to study the lipid dynamics of breast cancer cell,and proved the existence of the micro-domain in MCF-7cell membrane order.Finally,cholesterol were depleted by different concentrations of MβCD,and compared the diffusion coefficient and diffusion time of the MCF-7 cell membrane order and disorder under different concentrations of MβCD.The study showed that the distributions of the raft marker(CTXB)change obviously,indicate that the behavior of the membrane order phase become more fluidity,however,whereas the distributions of the non-raft markers(DilC18)remained similar as before the drug treatment showing their non-dependency on cholesterol.These results may be helpful to further study the role of membrane lipid in the processes of breast cancer cell migration and signal transduction,and then provide new ways for targeting lipid rafts breast cancer treatment.. |
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