Impact of deubiquitinating enzyme PSMD7 on bladder cancer proliferation and migration
Objective To investigate the effect of deubiquitinating enzyme PSMD7 expression on bladder cancer proliferation and migration.Methods The differential expression of PSMD7 in bladder cancer was analyzed by Timer database.In bladder cancer cell lines(UMUC3 and T24),regular interval short palindromic repeat clusters(CRISPR)/Cas9 technology was used to introduce short hairpin RNA(shRNA)to knock down PSMD7.The following groups were set up:control group and stable PSMD7 knockout experimental groups[UMUC3(UM-sh-1 group and UM-sh-2 group)and T24(T24-SH-1 group and T24-SH-2 group)].The effects of PSMD7 on the proliferation and migration of bladder cancer cells was investigated by cell counting kit-8(CCK-8)assay,plate clone formation assay and Transwell assay.Cell proliferation marker 5-acetylidene-2'-deoxyuridine(EDU)was detected by immunofluorescence assay to verify that PSMD7 knockdown inhibited the proliferation of bladder cancer cells.The independent sample t test was used for comparison between groups.Results The Timer database found that PSMD7 was highly expressed in a variety of cancers(P＜0.05),including bladder cancer,and the expression of PSMD7 in tumour tissues was higher than that in normal tissues,with a statistically significant difference[8.3(7.5-9.1)vs.3.1(2.2-4.1),P＜0.05].The CCK-8 results showed that the proliferative capacity of the cells was significantly decreased after knockdown of PSMD7 as compared with the control group[the absorbance of the control group was significantly higher than that of the knockdown group in the UMUC3 cell line(1.724±0.057 vs.0.879±0.040,t=20.83,P＜0.05)and(1.724±0.057 vs.0.935±0.065,t=15.62,P＜0.05);The absorbance of the control group was significantly higher than that of the knockdown group in the T24 cell line(2.329±0.084 vs.0.973±0.072,t=21.14,P＜0.05)and(2.329±0.084 vs.0.830±0.087,t=21.41,P＜0.05)].The results of the plate clone formation assay showed that cell proliferation ability was significantly decreased after PSMD7 knockdown[The number of colonies in the control group in UMUC3 cell line was significantly greater than that in the knockdown group(716.5±18.5 vs.508.0±21.0,t=7.45,P＜0.05)and(716.5±18.5 vs.396.5±9.5,t=15.39,P＜0.05);the number of colonies in the control group in T24 cell line was significantly greater than that in the knockdown group(687.0±33.0 vs.359.5±15.5,t=8.983,P＜0.05)and(687.0±33.0 vs.407.0±18.0,t=7.449,P＜0.05)].Transwell assay showed that the cell migration ability was significantly decreased after PSMD7 knockdown[The number of migrating cells in the control group in the UMUC3 cell line was signifi-cantly greater than that in the knockdown group(756.0±28.0)vs.(198.0±9.5),t=18.86,P＜0.05 and(756.0±28.0)cells vs.(146.5±11.5)cells,t=20.14,P＜0.05;the number of migrating cells in control group in T24 cell line was significantly greater than that in knockdown group(648.0±18.0)vs.(132.0±7.0),t=25.79,P＜0.05 and(648.0±18.0)vs.(159.0±13.0),t=21.21,P＜0.05].Immunofluorescence experiments showed that EDU was significantly higher in the control group than in the knockdown group[EDU was significantly higher in the control group than in the knockdown group in the UMUC3 cell line(0.873±0.054 vs.0.137±0.026,t=11.16,P＜0.05)and(0.873±0.054 vs.0.184±0.054,t=8.19,P＜0.05);EDU was significantly higher in the control group than in the knock-down group in the T24 cells(0.869±0.025 vs.0.156±0.013,t=25.02,P＜0.05)and(0.869± 0.025 vs.0.125±0.005,t=28.63,P＜0.05)].Conclusion PSMD7 expression is upregulated in bladder cancer,and PSMD7 can promote the proliferation and migration of bladder cancer cells.
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