P27 Tumor Suppressor Gene Transfection In Vitro Inhibition Of Human Tenon Fibroblast Cells

Objective:Glaucoma is a guoup of disease which has a common feature of optic atrophy and visual field defect,it has been arranged for the second place and the blinding process is irreversible.Currently the treatment of glaucoma filtration surgery is the most common surgical procedures. However,within 2 years after glaucoma filtering surgery, the failure rate is 15-25%.The main reason is the surgical area fibroblast proliferation, extracellular matrix synthesis,collagen deposition,and followed by subcon-junctival fibrosis occurs.The filtration path of surgery scar formation resulted the bleb. Dysfunction,Ultimately lead to the surgical failure.To suppress the formation of scar filtration road, it is often used the inhebit formation of scar drugs in filtering surgery,To delay or inhibit scar formation filtration road,to improve the success rate of surgery.The present study the anti-scarring drugs used are 5-fluorouracil,mitomycin,etc,their always have some inhibitory effects,but the inhibition is lacking the targeting. Although these drugs on cell proliferation have different degrees of inhibition,But because there have inevitable side effects,so clinical application is limited to a certain degree.P27 tumor suppressor gene is the cyclin-dependent kinase inhibitor, control of cell cycle from G1-S phase transition critical.This experiment is to discussion the cationic liposome-mediated P27 tumor suppressor gene in cultured human Tenon fibroblast inhibition in vitro. For clinical gene therapy scarring after glaucoma filtering surgery to provide a theoretical basis.Mcthods:Primary cultured human Tenon fibroblasts, immunohis-tochemical identification of the third generation used in the experiment, Using liposome-mediated method,the constructed PEGFP-N1P27 eukaryotic expression plasmid transfection in vitro cultured human Tenon fibroblasts, recombinant plasmid PEGFP-N1 as the empty vectou transfection group, cultured fibroblasts as a blank guoup.The cell cycle was observed by flow cytometry.P27 gene on the role of fibroblast by a methylthiazlyl tetrazolium (MTT)assay.Results:The cell cycle in P27 gene transfection group is almost in the G0-G1 phase,PEGFP-N1 transfected empty plasmid group and control group most of the cell cycle in S phase,P27 tumor suppressor gene transfection allows cell cycle arrest in G1 phase,P27 gene transfection significantly inhibited the proliferation of fibroblasts.Conclusion:The P27 gene transfection in vitro may inhibited the pro-liferation of fibroblasts on cultured human Tenon fibroblasts.