Experimental Study Of Effects Of P16-gene Transfer On Cell Growth Of The Human Bladder Carcinoma In Vitro

The main purpose of this study is to examined the effect of full length p16 gene transfer by replication-defective recombinant adenovirus on cell growth of the human bladder careinoma.Human embry oidney cell line 293 were used to generate recombinant adenovirus containing human p16 cDNA (Ad-p16). Ad-p16 was infected into the human bladder carcinoma cell line with no expression of p16 protein,T24.We tested the transfection efficiency of Ad-p16 using Ad-LacZ. The expression of p16 protein was evaluated by Western-blot analysis and Flow Cytometer. The effects of Ad-p16 infection on cell proliferation were evaluated by cell count and MIT assay. We also tested the effects of the expression of p16-gene on the expressions of some growth or apotosis-related proteins, such as AgNORs,PCNA and p53 protein.The experiment suggested:1 Higher transfection efficiency to T24 could be obtained at MOI of 50. 2. This Ad-p16 hasa high gene expression in T24 cells. 3 p16-gene expressioninhibited T24 cell proliferation through G1-S checkpoint. 4p16 protein has negtive effects on the expression of AgNORsand PCNA, but no effect on the expression of p53 protein.