Experimental Study On The Affection Of Exogenous Expression Of Activin Receptor ⅡA On Proliferation Of Human Lens Epithelial Cells

Objective：To investigate the affection of Exogenous expression of activin receptorⅡA on Proliferation of human lens epithelial cell line SRA01/04(HLECSRA01/04).Methods：(l) The plasmid ActRⅡA was digested, and then connected to theeukaryotic expression vector PC-DNA3.0.They will co-express a fusion proteinof c-myc, then the recombinant plasmid PC-DNA3.0of c-myc-ActRⅡA wassuccessfully constructed. LDMEM medium containing10%fetal calf serum(FBS) is used for culturing HLECs. The recombinant plasmid PC-DNA3.0c-myc-ActRⅡA and plasmid PC-DNA3.0can be transferred to the cells bycationic liposome mediating.48h after transfection, the cells were screened toestablish a stable cell line expressing ActRⅡA with G418of800ug/mlconcentration. The expression of ActRⅡA in HLECs can be detected withWestern-blot method.(2) The proliferation of stable transfected cell line can be detected bymethyl thiazolyl tetrazolium (MTT) colorimetry.Results：Higher expression of ActRⅡA of the experimental group can be detectedby Western-blot than control group. It shows that cell proliferation of theexperimental group was significantly inhibited, and the inhibition become moreand more obvious with time. There were absolutely significant differencesbetween the two groups (P<0.01).Conclusion:Exogenous transferred activin receptor ⅡA (ActRⅡA) has higherexpression in HLECs of experimental group. HLECs cell line stably expressing ActRⅡA was successfully constructed. ActRⅡA has significantly inhibited theproliferation of HLECs, and it will lay a theoretical basis for gene therapy ofafter cataract.