Objective      To determine the effect of membrane-cytoskelet on linker Ezrin in the invasion and migration of human renal carcinoma cells line 786-0.       Methods      With ezrin as the target gene

2 pairs of shRNA was designed and chemically synthesized

and then inserted into plasmid pGenesil-shRNA. The recombinant plasmid was transformed into DH5α. The recombinant plasmid was extracted in middle quantity and transfected into 786-0 cells line by Lipofectamine 2000. Ezrin expression in 786-0 cells transfe

cted by shRNA recombinant plasmid was measured by qRT-PCR and Western blotting. 786-0 cells were transfected by effective shRNA plasmid and cultured in 1640 media containing G418 (800 μg/ml). qRT-PCR and Western blotting were applied respectively to detect the Ezrin mRNA and protein expression to evaluate the blocking effect of shRNA-Ezrin. MTT assay and flow cytometry was used to further assess cell proliferation and apoptosis. Migration and invasion was determined by cell adhesion analysis and chemotactic motion assay.       Results      Our designed shRNA knocked down 93% Ezrin in 786-0 cells which had a stable expression of Ezrin small hairpin RNAs. After transfected with p-GeneSil-l-ezrin

the capability of 786-0 cells of proliferation

invasion and adhesion were decreased dramatically. Compared with the cells treated with shRNA-ezrin1

the percentage of G

0

/G

1

cells was significantly higher than those of untransfected cells and 786-0cells transfected with shRNA-hk（

P

<

0.01）

and proliferation index (PI) was decreased in the cells treated with shRNA-ezrin1 compared with those of untransfected cells and 786-0cells transfected with shRNA-hk（

P

<

0.01）. Heterotypic cell adhesion analysis showed that the numbers of adhesive cell in 786-0 cells transfected with shRNA-ezrin1 were less than the 786-0 cells untransfected and transfected with shRNA-hk (

P

<

0.01). Chemotactic motion was decreased in 786-0 cells transfected with shRNA-ezrin1.       Conclusion      Ezrin plays an important role in cell migration and invasion of human renal carcinoma cells

which may be regarded as a promising target for tumor gene therapy.