Expression of tripartite motif-containing protein 22 in prostate cancer and its impact on ferroptosis of prostate cancer cells
Objective To detect the expression levels of tripartite motif-containing protein 22(TRIM22)in prostate cancer tissues and cells,and to explore the effects of TRIM22 on the biological func-tions of prostate cancer cells.Methods The GEPIA database was used to analyze the expression level of the TRIM22 gene in prostate cancer tissues.A total of 16 male prostate cancer and adjacent tissue samples[age(64.45±11.34)years]were collected from Renmin Hospital of Wuhan University.The expression differences of TRIM22 mRNA and protein in prostate cancer and adjacent tissues,as well as in prostate cancer cell lines PC3,DU145,and normal prostate cell line RWPE-1,were detected using real-time quan-titative reverse transcription polymerase chain reaction(RT-qPCR)and Western blot techniques.Immuno-histochemical staining was used to detect the expression of TR1M22 protein in prostate cancer and adjacent tissues.Subsequently,TRIM22 overexpression plasmid and its control were transfected into PC3 and DU145 cells.The cells were divided into PC3 experimental group,PC3 control group,DU145 experimental group,and DU 145 control group.The transfection efficiency was verified by RT-qPCR and Western blot.The effects of TRIM22 overexpression on cell proliferation were evaluated using colony formation and EdU assays.The influence of overexpression on the expression of key proteins related to ferroptosis was verified by Western blot.Inter-group comparisons of continuous data were performed using the t-test.Results The GEPIA database shows that the expression level of the TRIM22 gene is decreased in prostate cancer tissues(P＜0.05).RT-qPCR and Western blotting results indicate that the relative expression levels of TRIM22 mRNA and protein in adjacent normal tissues are higher than in prostate cancer tissues(mRNA:1.42±0.04 vs.1.01±0.06,t=5.590,P＜0.05;protein:156.67±5.54 vs.97.67±3.76,t=8.806,P＜0.05).The relative expression levels of TRIM22 in PC3 and DU145 cells are lower than in RWPE-1 cells(mRNA:0.51±0.05 vs.1.03±0.05,0.61±0.05 vs.1.03±0.05,t=7.320,5.080,P＜0.05;pro-tein:56.00±4.04 vs.104.05±5.19,67.33±3.76 vs.104.05±5.19,t=7.292,5.719,P＜0.05).In the experimental PC3 and DU145 cells,the relative expression levels of TRIM22 mRNA and protein are higher than those in the control group(mRNA:7.24±0.16 vs.1.03±0.07,5.93±0.14 vs.0.98±0.07,t=36.224,46.048,P＜0.01;protein:552.33±14.01 vs.100.67±8.74,435.67±14.05 vs.100.33±12.53,t=47.376,30.886,P＜0.01).The results of the cell colony formation assay show that the number of colonies formed by experimental PC3 and DU 145 cells is lower than that of the control group[(34.33±7.02)cells vs.(61.33±6.56)cells,(30.33±4.04)cells vs.(55.67±6.51)cells,t=4.807,5.729,P＜0.05].In the EdU assay,the proliferation activity of experimental PC3 and DU 145 cells is lower than that of the control group[(24.33±4.33)％vs.(81.67±3.76)％,(20.67±4.41)％vs.(79.33±6.24)％,t=9.997,8.568,P＜0.05].Western blot results suggest that the expression of ferropto-sis-related proteins GPX4 and SLC7A11 in the experimental PC3 and DU145 cells does not show significant changes compared to the control group(GPX4:98.67±10.01 vs.99.67±7.02,99.33±9.61 vs.101.33±9.02,t=0.142,0.263,P＞0.05;SLC7A11:98.67±6.11 vs.102.67±7.51,100.67±8.02 vs.99.00±8.54,t=0.716,0.905,P＞0.05).However,the expression of NRF2/HO-1 pathway proteins in the experimental PC3 and DU145 cells is significantly reduced compared to the control group(NRF2:28.67±6.03 vs.104.33±10.79,46.33±7.51 vs.101.33±6.66,t=10.607,9.695,P＜0.05;HO-1:13.33±7.09 vs.102.67±7.51,25.33±7.52 vs.103.33±8.02,t=14.982,12.299,P＜0.05).Conclusion TRIM22 is abnormally low expressed in prostate cancer tissues and cells,suggesting that it may function as a tumor suppressor gene inhibiting the occurrence and development of prostate cancer.
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