The effect of cyclin dependent kinase 5 on the growth,invasion,and metastasis of castrated-resistant prostate cancer cells
Objective To study the effect of cyclin dependent kinase 5(CDK5)on the growth,invasion,and metastasis of castrated-resistant prostate cancer(CRPC)cells.Methods CDK5 small molecule inhibitor 20-223 was added to cells at four levels of concentration,with 0.00 μmol/L for control group,2.00 μmol/L for 2.00 μmol/L group,4.00 μmol/L for 4.00 μmol/L group and 8.00 μmol/L for 8.00 μmol/L group,with 100 stains in each group.CDK5 small interfering RNA(siRNA)was used to interfere with the expression of CDK5 in DU145 cells,and CDK5 inhibitor 20-223 inhibited the activity of CDK5 in DU145 cells;Cell growth activity was measured using methyl-thiazole-tetrazolium(MTT),and cell proliferation ability was measured using monoclonal formation assay;Cell apoptosis was measured using Annexin V-fluorescein isothio-cyanate(Annexin V-FITC)and propidium iodide(PI)double staining methods;The scratch test and Transwell test were used to determine the migration ability of cells;Western Blot was used to determine the expression of Myc oncogenic gene(c-Myc),SRY-box transcription factor 4(SOX4),and invasion and metastasis related proteins.Comparatively analysis of CDK5 inhibitor 20-223 on CRPC cell DU145 growth,apoptosis,invasive metastasis ability,as well as the expression of c-Myc,SOX4,and epithelial mesenchymal transition(EMT)-related proteins in the four groups of cells.Results The cell growth inhibition rates of the 2.00 µmol/L group,4.00 μmol/L group,and 8.00 µmol/L group were higher than those of the control group at 24,48,and 72 h after medication,with a statistically significant difference(P<0.01).CRPC cell DU145 growth viability decreased with increasing concentrations of CDK5 inhibitors and their growth was significantly inhibited.The incidence of cell apoptosis of Q1 quadrant,Q2 quadrant,Q3 quadrant,and Q4 quadrant in the 2.00 µmol/L group,4.00 μmol/L group,and 8.00 µmol/L group were higher than those in the control group at 48 h after medication,with a statistically significant difference(P<0.01).The migration rate in the 4.00 μmol/L group was 2.28%at 48 h after medication,which was lower than 2.37%in the control group,with a statistically significant difference(P<0.01).After 2 days of treatment,the expression levels of c-Myc,SOX4,Vimentin,and Zeb1 proteins in the 2.00 µmol/L group,4.00 μmol/L group,and 8.00 µmol/L group were lower than those of the control group,while the expression of E-cadherin was higher than that of the control group.The expression of CDK5,c-Myc,and SOX4 proteins in CDK5 gene silenced tissues was significantly lower than that in the control group and NC group(P<0.05).Conclusion The decreased expression and activity of CDK5 in CRPC can inhibit the malignant proliferation and infiltration of DU145 cells to a certain extent,and promote their apoptosis.CDK5 has a strong potential and belongs to a castration-resistant drug.When its activity is inhibited,the levels of c-Myc,SOX4 and other proteins are significantly decreased,and then the levels of E-cadherin,Vimentin,Zeb1 and other proteins are up-regulated,thus weakening the sensitivity of castration-resistant drugs.
Cyclin dependent kinase 5Castrated-resistant prostate cancerInvasion and metastasisApoptosis
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NSTL
万方数据
维普
