Influence Of △NP63 Silence On Proliferation And Its Molecular Mechanism

Objective : To observe the influence ofâ–³Np63 gene on the proliferation of Transitional cell carcinoma of bladder tumor cells, and initially analyze the possible mechanism of it.Methods: Transitional cell carcinoma of bladder 5637 cells were transfected withâ–³Np63 specific oligonucleotides(â–³Np63-shRNA) and Control-shRNA. 5637 cells were divided into three different treatment groups: blank control group, Control-shRNA group andâ–³N p63-shRNA group. The fluorescence was used to confirm the success of transfection in 5637 cells. The cell proliferation was assessed by WST-1 assay, and the changes in cell cycle was evaluated by flow cytometry, Reverse transcriptase PCR were used to measureâ–³Np63, p27^kip1 and p57^kip2 expression at mRNA level respectively, western bolt were used to measure theâ–³Np63, p27^kip1 and p57^kip2 expression at protein level respectively.Results: The 5637 cells was successfully transfected byâ–³Np63-shRNA expression plasmid and confirmed by fluorescence microscope. The proliferate inhibition rate of 5637 cells inâ–³Np63-shRNA group at 24h, 48h and 72 were 12.3±5.8%,42.7±6.7% and 51.9±7.1% respectively,it was markedly inhibited of 5637 cells was increased transfected byâ–³Np63-shRNA(P<0.05). The G₀/G_l cycle was increased and S cycle was decreased inâ–³Np63-shRNA group, The specific shRNA targeted toâ–³Np63 gene could down regulate the expression ofâ–³Np63 and up regulate the expression of p27^kip1, p57^kip2 at mRNA and protein level.Conclusion: The 5637 cells could been successfully transfected withâ–³Np63-shRNA expression plasmid,â–³Np63 can effectively inhibit the expression ofâ–³Np63 protein and mRNA andup regulate the expression of p27^kip1 and p57^kip2. Theâ–³Np63-shRNA could effectively induce the growth inhibit in the way of up regulate the expression of p27^kip1 and p57^kip2.