Study On Cytoskeleton Regulation By Human MiR-941

miR-941 is a human-specific microRNA discovered in our previous work.It may be related to human evolution,yet its cell function is unknown.Through the establishment of miR-941 overexpression stable cell lines,this thesis investigated the potential functions of miR-941 at the cellular and molecular levels.In the 293 T cells stably overexpressing miR-941,no significant effect on the proliferation and migration of 293 T cells was detected by EdU method and Transwell assay,while cytometry revealed that miR-941 significantly promoted cell necrosis.Immunofluorescence staining of microfilaments and microtubules revealed that miR-941 reduced the number of 293 T cells with the centrosome-rich microtubule pattern,cell protrusions became thin and the area and distribution of focal adhesions in these cells was altered.The traits of microtubule and microfilament influenced by miR-941 overexpression was further investigated in Hela cells.No obvious effect was found in microtubule distribution,but the microfilament skeleton at the edge of cells was more obvious when miR-941 was overexpressed.Cell cycle detection showed that there were more S phase cells was observed;when treated with the microtubule depolymerizing drug Vincristine,the proliferation of Hela cells decreased,but miR-941 overexpressing cells were significantly less affected than the control;under the action of microfilament polymerization drug Jasplakinolide,microfilaments in miR-941 overexpressed cells showed less change,whereas the depolymerization of microfilaments affected by LatrunculinA were more serious.QRT-PCR showed that the expression levels of PRKAG2 and other cytoskeleton-related protein genes were affected by miR-941 overexpression,in which PRKAG2 and KLHL2 were significantly up-regulated and down-regulated,respectively,and their regulatory mechanisms remain further study.These findings suggested that the occurrence of miR-941 may promote the dynamics of the cytoskeleton,leading to cytoskeletal plasticity in human cells in response to external influences.