Ubiquitin specific protease 41 is upregulated in breast cancer and and its correlation with malignant biological behaviors
Objective To explore the potential value of ubiquitin specific protease 41(USP41)as novel diagnostic/prognostic biomarker for breast cancer and determine the correlation between USP41 and malignant biological behaviors.Methods Transcriptome data and clinical information of 1086 breast canc-er samples from The Cancer Genome Atlas(TCGA)database and 3273 breast cancer samples from the GSE96058 dataset in the Gene Expression Omnibus(GEO)database were obtained.Gene expression anal-ysis,prognosis analysis,functional enrichment analysis,and immune infiltration analysis were conducted using R 4.1.2.Two specific small interfering RNAs(siRNAs)were transfected into the breast cancer cell line MCF-7,and Western blotting was used to detect the protein expression levels of USP41 before and after transfection.Colony formation assay,cell counting kit-8(CCK-8)assay,wound healing assay,and Tran-swell assay were used to assess the proliferation,invasion,and migration abilities of breast cancer cells af-ter transfection.Cox regression was used to compare prognostic differences.The comparison of relative grayscale values was performed by one-way analysis of variance and Tukey HSD post hoc test.The compari-son of cell colony number was performed by Welch's analysis of variance and Games-Howell post hoc test.The comparison of absorbance values and relative widths at the same time was performed by two-factor re-peated measures data analysis of variance and multiple pair comparisons.The comparison of cell migration number was performed by Kruskal-Wallis test and Dunn's post hoc test.Single-factor and multi-factor Cox regression was used to determine independent prognostic risk factors.Correlation analysis was performed u-sing Spearman analysis.Results USP41 was highly expressed in 10 types of cancer tissues,and its diag-nostic efficacy as a diagnostic marker for breast cancer was moderate(area under the curve=0.717).High expression of USP41 was associated with poorer overall survival in breast cancer patients[P＜0.01,hazard ratio(HR)=1.93,95％confidence interval(CI):1.40-2.66]and disease-free survival(P＜0.01,HR=2.10,95％CI:1.37-3.23].Cox regression analysis indicated that USP41 was an independent risk factor for breast cancer(P＜0.01,HR=1.406,95％CI:1.13-1.75).Gene set enrichment analysis revealed that high expression of USP41 was associated with functional activities related to dopaminergic neurogene-sis,dopamine neurotransmitter release cycle,and insulin uptake.Immune infiltration analysis showed that high expression of USP41 was significantly positively correlated with increased immune cell infiltration and expression of multiple immune checkpoints.The protein expression levels of USP41 in siRNA1 and siRNA2 groups(3.88±0.27,4.13±0.12)were significantly lower than those in the control group(6.03±0.40,F=194.405,P＜0.01).The number of colonies in siRNA1 and siRNA2 groups[(20.75±6.13),(25.50±8.89)]was significantly less than that in the control group(76.75±17.23)(F=16.720,P＜0.01).The absorbance values at 24 h for siRNA1 and siRNA2 groups(0.43±0.01,0.44±0.01)were significantly lower than those for the control group(0.51±0.04)(F=11.933,P＜0.05).The invasion widths at 24 h for siRNA1 and siRNA2 groups(2.53±0.12,2.67±0.12)were significantly greater than those for the control group(1.70±0.10)(F=67.364,P＜0.01).The number of migrating cells at 24 h for siRNA1 group(190.50±19.09)was significantly less than that for the control group(351.25±15.88,F=-2.755,P＜0.05).Conclusion High expression of USP41 may be correlated with poor prognosis and enhance malignant biological behaviors of breast cancer.
Ubiquitin specific protease 41Breast cancerBioinformaticsPrognosisBio-logical behavior
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