Researches On The Mechanical Characteristics Of The Kidney Tumor Cell Of Mice

Cell Traction force(CTF) is the force exerted on the extracellular substrate by the cell. It isformed by the interaction of actomyosin and the polymerization of actin. The CTF is regulatedby α-SMA and exert the extracellular substrate via focal adhesion. CTF has close relationswith many biological processes as an important physical parameters. It can affect significantlyto cell’s attach, movement, morphology and signal transduction etc. We attempt to discoverthe change mechanism of CTF of mice kidney during the cancelation through the study thechange of mechanical properties of mice kidney tumor cells during the cancelation.Objective: Through the observing the change of the CTF and the α-SMA of mice kidneytumor cells during the induced differentiation and comparing the normal mice kidney cells,discover the regular patter of the change of CTF and α-SMA in the process. Thereby explorethe change mechanism of CTF and α-SMA in the process of growth, development andpathology and do the some exploratory research on basic biology.Method and conclusion:1. Cell culture. Use the digestion method to obtain the normal cells and tumor cells of micekidney, and then put it in the CO2incubator to cultivate. Change the medium and remove non-attach cells after3-4days. Observe the cell growth morphology with inverted microscope,When the most of cells get good colony make the glass sample and keep on culture for use.2. Stain microfilament with Rhodamine Phalloidin. Stain the microfilament withRhodamine Phalloidin as which combine specially with microfilament. And than, comparethe difference of the distribution of microfilament between the mice kidney normal cells andtumor cells. The result shows that:the distribution of microfilament of mice kidney tumor cellsare disorder compar with normal kidney, they have changed much. This prove that the cells’structure of skeleton have changed dramatically after cancelation, which adapted to thephysiological characteristics of tumor cells.3. Measure CTF with CTFM method. CTFM is the technology which basic on surfacedeformation of cellular elastic substrate to calculate CTF. According to the different size of thecells make different substrate on hardness and add2%fluorescent microsphere for to measurethe surface deformation of cellular elastic substrate easily. Seed cells in the elastic substrate surface, the substrate will distort after cells attach on the surface. At first use the ordinary lightof fluorescence microscope to take photo, and then use the fluorescent light to take the substrateimages, and then use1drop1M NaOH to kill the cells, make them de-attach from the surfaceof the substrate and elastic substrate will restore prototype, and then use the fluorescent light totake the second substrate images. Run the software in MATLAB7.0to calculate the CTF. Usethe same method to get the CTF of mice kidney tumor cells before and after induceddifferentiation.The result shows that CTF became to bigger after induced differentiation than before, andthe average increased from281Pa to550Pa increased about100%. The projective areasincreased, the average of projective areas increased from695um2to955um2, increased about37%. The maximum of CTF of the mice kidney tumor cells increased from1200Pa to1700Pa.4. Measure the α-SMA protein content with western blot method. Use mouse anti α-SMAimmunoglobulin as first antibody and use HRP sheep anti mouse immunoglobulin as secondantibody, use ECL Kit to development. The result shows that α-SMA in kidney tumor cellsincreased after differentiation, the content close to the normal cells. This shows cinnamic acidhave good induced effect for mice kidney tumor cells.