| Objective:To investigate the inhibition of siRNA-YAP1 to epithelial-mesenchymal transition（EMT）of human lens epithelial cells（LECs）induced by TGF-β2.Methods:Human LECs line（HLEB-3）was cultured and divided into control group and TGF-β2induced group,and 10 ng/ml TGF-β2was used to treat the cells in the TGF-β2induced group.The expressions of E-cadherin and Vimentin proteins were detected in different groups by Western blot;Real-time fluorescence quantitative PCR（q-PCR）,immunofluorescence assay,and Western blot were used to detect the expression of Yes-associated protein 1（YAP1）in cells,respectively.Cultured cells were divided into siRNA-NC group（NC）,siRNA-YAP1 group（si-YAP1）,siRNA-NC TGF-β2group（NC+T）and siRNA-YAP1 TGF-β2group（si-YAP1+T）,transfected cells with siRNA-NC plasmid and the best siRNA-YAP1 plasmid respectively.The relative expression of YAP1 m RNA and protein in various groups were detected by q-PCR,immunofluorescence assay and Western blot,respectively.Detection of EMT marker protein in cells by Western blot and immunofluorescence assay.Results:The E-cadherin protein（0.797±0.110）in the TGF-β2 group was lower than that in the control group（1.240±0.110）,and the Vimentin protein（1.18±0.118）in the TGF-β2 group was higher than the control group（0.830±0.104）;The YAP1 m RNA（2.200±0.193）in TGF-β2the group was higher than that in the control group（1.136±0.123）.The YAP1 protein（1.203±0.121）in the TGF-β2group was higher than that in the control group（0.967±0.025）.The YAP1 protein fluorescence in the TGF-β2group was lower than control group;the differences were statistically significant（P<0.05）.In the si-YAP1 group,the YAP1 protein（0.303±0.057）and YAP1 m RNA（0.576±0.108）were lower than YAP1 proteinin（1.327±0.021）and YAP1 m RNA（1.156±0.175）in the NC group.Compared with the NC+T group,the YAP1 fluorescence in the si-YAP1+T group was significantly reduced;in the si-YAP1+T group,the YAP1 protein（0.320±0.104）and YAP1 m RNA（0.572±0.022）were lower than YAP1 protein（1.453±0.021）and YAP1 m RNA（2.048±0.329）in the NC+T group;the differences were statistically significant（P<0.05）;YAP1 protein fluorescence expression showed the same trend in different groups.E-cadherin protein expression in NC+T group（0.560±0.029）was lower than NC group（0.817±0.037）,si-YAP1+T（0.744±0.092）was higher than NC+T group（0.560±0.029）,si-YAP1 group（0.987±0.041）was higher than the NC group（0.817±0.037）,and the differences were statistically significant（P<0.05）.The fluorescence expression of E-cadherin protein showed the same trend in different groups.Vimentin protein expression in NC+T group（1.005±0.032）was higher than NC group（0.788±0.018）,si-YAP1+T（0.791±0.021）was lower than NC+T group（1.005±0.032）,and the differences were statistically significant.Significance（P<0.05）;vimentin protein expression in the si-YAP1group（0.745±0.017）and NC group（0.788±0.018）showed no significant difference（P>0.05）.Vimentin protein fluorescence expression showed the same trend in different groups.Conclusions:YAP1 participates in the EMT of human LECs induced by TGF-β2,and siRNA-YAP1 can suppress the EMT process. |
