Research On Mechanical Property Of Hepatoma Cells(Hepa1-6)of Mous

Cell traction forces (CTFs) are generated by actomyosin interactions and actin polymerization and regulated by intracellular proteins such as alpha-smooth muscle actin (α-SMA) and soluble factors such as transforming growth factor-β (TGF-β). Once transmitted to the extracellular matrix (ECM) through stress fibers via focal adhesions, which are assemblies of ECM proteins.. Cell traction forces (CTFs) are crucial to many biological processes such as inflammation, wound healing, angiogenesis, and metastasis. Cell traction force is capable of facilitating cell locomotion, feeling the environment of extracellular matrix, maintaining physiological forms of cells and physiological characteristics of cells, etc. In short, it is influencing the growth, differentiation, pathology and apoptosis processes of cells. Hence, researches on cell traction force enable people to have a more comprehensive and profound understanding on cells and their changing mechanisms in the process of growing up. This experiment, through testing changes of cell traction force before and after the happening of mouse liver cell cancer, has studied the changing mechanism of cell traction force before and after the happening of liver cell cancer with a view to discovering the interrelation between cell traction force and cancer and illuminating the influencing mechanism.Objectives:to identify the changing mechanism of traction force of mouse Hepal-6cells and have a more profound understanding on the changing mechanism of mouse liver cell cancer and a more comprehensive knowledge on the biological properties of mouse Hepal-6cells through measuring the cell traction forces of mouse hepatic cells and Hepal-6cells.1. Preparation of cells. Attain mouse liver cells and mouse Hepal-6cells with digestion method; make cell slides or resuspend them when most cells have formed desirable colonies.2. Specific staining on microfilaments with phalloidin. Conduct a specific staining on microfilaments by making advantage of phalloidin as a specific drug so as to compare the changes of microfilament distribution features of mouse hepatic cells and mouse Hepal-6cells. The result indicated that:the microfilament distribution of Hepal-6cells was disordered and scattered in structure and the change was more obvious than that of mouse hepatic cells, which means the skeleton structure of cells had changed after the happening of cell canceration and that was compatible with physiological characteristics of cancer cells. 3. Testing of cell traction force、area of cell with cell traction force microscopy, to extract and analyze the information implied in the photos with MATLAB7.0specific self-compiled program. The result indicated that:compared with that of mouse hepatic cells, the cell traction force of mouse Hepal-6cells was less by about53%; the area of cell was less by about15%. The results suggest that the relationship between cell traction force and carcinogenesis is closely related.4. Testing on expression quantity of α-SMA in mouse Hepal-6cells with immunofluorescence and western blot method, Mark the expression intensity of α-SMA on the single cell level. The result indicated that:compared with that of mouse hepatic cells, the expression quantity of internal α-SMA of mouse Hepal-6cells has gone through obvious changes; on the single cell level, compared with that of mouse hepatic cells, the protein content of mouse Hepal-6cell was less by about52%; on the overall level, it was less by about47%. The results suggest that the α-SMA protein be related to the process of canceration and cell traction force。...