Abstract:

To explore the influence of   human thermal transient receptor potential 1 (TRP1)  gene transfection in expression of P27 protein in corneal endothelial cells   in cats and the effect of termal TRP1 gene in  proliferation cycles of corneal endothelial cells and to provide theoretical foundation for the application of the gene in human corneal endothelial cells.Methods 20 healthy full-term kittens were randomly divided into experiment group and control group.The corneal endothelial cells were collected after anesthesia and cultured.The cells in experiment  group were transfected with  thermal TRP1 gene mediated by liposome.The cells in control group were treated with 5.0 μL PBS instead of the expression vector into medium.The mRNA  expression  of thermal TRP1 gene was detected by RT-PCR.The expression of P27  protein  in cat corneal endothelial cells was observed by Western blotting.The influence of target gene transfection in the activities of cell division and regeneration 96 h after culture was observed.Results The inflow of calcium ions in the cultured cells  was increased significantly after heating.The RT-PCR  results of thermal TRP1 gene transfection showed that the strip density in experiment group was higher than that  in control group (t= 2.21,P= 0.037).The expression of  P27 protein in experiment group was significantly higher than that in  control group (t=2.53,P= 0.021).The differences of mitotic index and the proportion of the  cells at G1 phase  between experiment group and control group were statistically significant (χ2=3.26,χ2=4.18,P<0.05).Conclusion The human thermal  TRP1  gene transfection in cat corneal endothelial cells can increase the expression of  P27 protein and inhibit the proliferation of corneal endothelial cells.

Key words: thermal transient receptor potential1;gene transfection;corneal endothelial cells;P27 protein