The Regulating Effect Of Anillin On Cytoskeletal Protein And Cellular Mechanical Properties

Anillin is an important scaffolding protein and regulator in cells that interacts with various proteins such as actin fibers(F-actin),myosin II,RhoA and Septins throughout the cell cycle.It is necessary to maintain a proper balance of Anillin in cells.Overexpression of Anillin increases the migratory capabilities of cancer cells or leads to Focal segmental glomerulosclerosis(FSGS),while too little or no expression of Anillin will disrupt the he integrity of the contractile ring and cell membrane of dividing cells.In the meantime,mechanical properties such as cell surface elastic modulus are important indicators for distinguishing cancer cells.Therefore,studying the regulatory role of Anillin in cytoskeletal proteins and mechanical properties can provide valuable references for the clinical diagnosis and treatment of cancer and other diseases.This paper focuses on the effects of the deficiency of Anillin oncytoskeletal proteins and cellular mechanical properties.A Hela cell strain which can stably knock down Anillin RNA was obtained after screening by RNA interference technology,and then the mCherry-myosin II A fluorescent protein was transferred into cells.Using fluorescence recovery after photobleaching(FRAP)and linear fluorescence intensity analysis to measure the distribution and motion velocity of intracellular myosin II A along the long axis of cells,respectively;Fluorescence staining for detecting the distribution of actin fibers;Measure the surface elastic modulus at different positions of the cells and the membrane breakthrough force in the long-axis edge region by using atomic force microscope(AFM).The results are as follows:(1)The lack of Anillin leads to a decrease in the distribution of the long-axis end of myosin II and a slower motion velocity;(2)actin fibers aggregated at the edge of the cell but not regularly arranged,and the actin fibers in the cell body are not distributed along the long axis;(3)The surface elastic modulus of Anillin knockdown cells increased significantly,but there was no significant difference in the membrane breakthrough force at the long axis edge.The results of this paper show that: Anillin mainly regional regulates intracellular cells by acting on the skeletal structure and distribution of the cell edges.This result corresponds to the distribution of the cell surface elastic modulus,but does not significantly affect the membrane breakthrough force.Anillin affects physiological activities such as cell-cell interaction and polarity establishment by interacting with cytoskeletal proteins and cell surface elastic modulus.