Effect of ubiquitin carboxyl-terminal hydrolase 37 on the proliferation of SW1990 pancreatic cancer cells
Objective To observe the role of ubiquitin carboxyl-terminal hydrolase 37(UCH37)in regulating cell division cycle protein 25A(CDC25A)in SW1990 pancreatic cancer cells and to explore its effect on the proliferation of SW1990 cells.Methods The normal human pancreatic HPDE cells and pancreatic cancer SW1990 cells in logarithmic growth phase were harvested,and the relative expressions of UCH37 mRNA and protein were detected by real-time fluorescence quantitative PCR and Western blot.The SW1990 cells were transfected with sh-UCH37 lentivirus(UCH37 knockdown group)and UCH37-NC lentivirus(UCH37 blank group)for 48 h.The relative expressions of UCH37 and CDC25A mRNA and protein were detected by real-time fluorescence quantitative PCR and Western blot.The cell colony formation assay was used to detect the number of colony formation.The CCK-8 method was used to detect the optical density(OD)values of cell proliferation after culture for 24,48,72 and 96 h in two groups.Results(1)The relative expressions of UCH37 mRNA and protein were higher in SW1990 cells(3.338±0.137,0.956±0.043)than those in HPDE cells(0.968±0.040,0.056±0.003)(t=26.710,P＜0.001;t=36.080,P＜0.001).(2)After culture for 48 h,the relative expressions of UCH37 mRNA and protein were lower in UCH37 knockdown group(0.337±0.017,0.424±0.010)than those in UCH37 blank group(1.017±0.052,1.163±0.132)(t=21.810,P＜0.001;t=9.160,P＜0.001),and the number of colony formation was less in UCH37 knockdown group(27.0±7.9)than that in UCH37 blank group(125.3±14.1)(t=10.860,P＜0.001).The OD values after culture for 48,72 and 96 h were lower in UCH37 knockdown(0.433±0.016,0.563±0.004,0.897±0.046)than those in UCH37 blank group(0.531±0.014,0.826±0.017,1.859±0.126)(t=7.420,P=0.002;t=24.750,P＜0.001;t=12.300,P＜0.001),and there was no significant difference in the OD value after culture for 24 h between two groups(P＞0.05).(3)The relative expression of CDC25A protein was lower in UCH37 knockdown group(0.501±0.026)than that in UCH37 blank group(1.174± 0.038)(t=23.090,P＜0.001),while the relative expression of CDC25A mRNA showed no significant difference between two groups(P＞0.05).Conclusion The deubiquitinating enzyme UCH37 can regulate the cell cycle progression in pancreatic cancer cells through CDC25A,and to knockdown UCH37 expression can inhibit the proliferation of pancreatic cancer cells.
pancreatic cancerubiquitin carboxyl-terminal hydrolase 37cell division cycle protein 25A
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