| Background:Breast cancer is the second leading cause of cancer death in the world,of which recurrence and metastasis are the main causes of death.Triple-negative breast cancer(TNBC)has high histological grade,advanced stage,high rate of distant metastasis,and high lymphocyte infiltration density.OTUD6B is a member of the ovarian tumorassociated protease(OTU)family,which is involved in almost all physiological activities including protein-specific degradation,signal transduction and protein localization,and is closely related to the occurrence and development of various tumors.In this study,we comprehensively analyzed the role of the deubiquitinating enzyme OTUD6B in breast cancer(BRCA)by using data from a set of public databases.Secondly,combined with the target substrate of OTUD6B Forkhead box 3a(FOXO3a),the mechanism of action of the two in triple-negative breast cancer was discussed.Methods:We firstly downloaded the transcriptome sequencing data of BRCA and BRCA subgroups from The Cancer Genome Atlas(TCGA)and Gene Expression Omnibus(GEO)database,respectively.Then we analyzed the OTUD6B mRNA expression and further demonstrated the protein expression by Human Protein Atlas(HPA).Among them,R package was used to analyze the correlation between OTUD6B and breast cancer stage.ROC diagnostic curve,time dependent survival ROC curve and nomogram model were subsequently established.The cBioPortal and MethSurv were carried out to analyze the gene changes,mutation and DNA methylation of OTUD6B in breast cancer,and their effects on clinical prognosis.R software was then used to analyze data from TISIDB database in order to explore the correlation between OTUD6B and immune cell infiltration,immune cell biomarkers and immune checkpoints.And Gene Ontology(GO)analysis and Kyoto Encyclopedia of genes and genomes(KEGG)analysis were conducted to study the function of OTUD6B gene.What is more,for TNBC,First,we verified whether ubiquitination was involved in the regulation of FOXO3a levels in TNBC cells and whether the OTU family was involved in the regulation of FOXO3a levels.Secondly,qPCR was used to detect the expression levels of OTUD6B in TNBC patients’ cancer and adjacent tissues,CCK-8 and Transwell assays were used to detect cell proliferation and migration,and co-immunoprecipitation experiments were used to verify the interaction between FOXO3a and OTUD6B.Results:We found the OTUD6B is highly expressed in 23 cancers and OTUD6B is significantly correlated with molecular typing,estrogen,progesterone,and histological types of breast cancer.Patients with high expression of OTUD6B have a poor prognosis.OTUD6B helps to distinguish cancer from normal tissue(AUC=0.603).The area of 1-year,3-year and 5-year survival ROC was higher than 0.5.The change rate of OTUD6B gene was 13%,and the somatic mutation frequency was 0.1%.Of the 14 DNA methylated CpG sites,4 were associated with the prognosis of BRCA.The expression of OTUD6B was positively correlated or negatively correlated with 17 kinds of immune cells in breast cancer,and positively correlated with immunophenotyping cytotoxic T lymphocyte antigen 4(CTLA4)and programmed cell death protein ligand 1(PD-L1).KEGG pathway enrichment analysis revealed that OTUD6B was enriched in RNA transport and cell cycle pathways.In TNBC cells,OTUD6B may be a negative regulator of FOXO3a.The results of cell function experiments showed that interfering with OTUD6B significantly inhibited the proliferation and migration of TNBC cells.Secondly,Western blot,qPC and CoIP detection showed that overexpression of OTUD6B in TNBC cells significantly enhanced the ubiquitination of FOXO3a and down-regulated the level of FOX03a protein,but had no effect on its mRNA level.Given that MDM2 plays an important role in the ubiquitination and degradation of FOXO3a,we selected MDM2 as a candidate for study.Western blot showed that overexpression of OTUD6B significantly enhanced the protein level of MDM2 in TNBC cells,but had no effect on its mRNA level.Through coimmunoprecipitation experiments,it was found that OTUD6B can bind to MDM2 and inhibit the ubiquitination and degradation of MDM2.Conclusions:Overall,OTUD6B has good prognostic value and may be involved in the immune regulation of breast cancer,as well as a potential therapeutic target for patients with breast cancer.In addition,at the level of cytological function,OTUD6B may stabilize the level of MDM2 by binding and deubiquitinating MDM2 in TNBC cells,thereby enhancing the ubiquitination and degradation of FOXO3a by MDM2,downregulating its stability and activity,which results in occurrence,invasion and metastasis of triple-negative breast cancer.Background:Adenoid cystic carcinoma(ACC)of the breast is a rare type of breast cancer with unknown oncological behavior.This study retrospectively analyzed the clinical manifestations,imaging features and clinicopathological features of breast ACC,explored its prognostic factors,and constructed a prognostic nomogram in order to accurately predict the overall survival(OS)of breast ACC patients.Methods:Based on the Surveillance,Epidemiology,and End Results(SEER)and the Cancer Hospital of the Chinese Academy of Medical Sciences data,we screened out 420 patients with pathologically diagnosed breast ACC for their imaging features,clinical features Information on pathology and treatment.The patients in SEER database were divided into training set(n=285)and validation set(n=121)according to 7:3.Based on R software,the clinicopathological characteristics of patients in SEER database and our hospital database were compared.At the same time,univariate and multivariate Cox regression model were used in the training set to evaluate the prognostic factors,and the nomogram prognostic model was further constructed.Finally,the performance and benefit of the nomogram were evaluated using the C index,ROC curve(Receiver operating characteristic curve),calibration curve,and DCA curve(Decision curve analysis)in the training set and validation set.Results:We screened 406 patients with adenoid cystic carcinoma of the breast who met the inclusion and exclusion criteria(285 in the training set and 121 in the validation set)from the SEER database.A total of 14 patients were included in our hospital.Our data suggest that the most common clinical manifestation of breast ACC is a palpable mass(85.7%),and the imaging features of Color Doppler ultrasonography and mammography in all our patients were nonspecific.ACC was suspected by fine needle aspiration cytology(FNAC)in 6 patients(42.9%)in our hospital and confirmed by postoperative histology and immunohistochemistry.Compared with the SEER and the data of our hospital,the proportion of breast-conserving patients in our hospital 4(28.6%)was lower than that in the SEER database 273(67.2%).At the same time,the proportion of modified radical mastectomy for breast cancer in our hospital 4(28.6%)was significantly higher than that in SEER database 52(12.8%).Although Her-2 status and molecular subtype were statistically different between the two data,it may be due to the larger proportion of SEER database before 2010.For Her-2 status,the proportion of patients before 2010 was 227(55.9%),which was significantly higher than the 177(43.6%)of Her-2-negative cases and 2(0.5%)of positive cases.Significant factors affecting the prognosis,including age,histological grade and N stage were taken into the nomogram based on univariate and multivariate Cox regression analysis.The C-index of the nomogram in the training set was 0.738,while the areas under the ROC curve of the training set for predicting 3,5,and 10year overall survival were 0.749,0.782,and 0.80,respectively.the c-index of the validation set was 0.803 and the areas under the ROC curve for the validation set to predict 3,5,and 10-year overall survival were 0.780,0.833,and 0.779,respectively.Such results suggest that the nomogram has a good prognostic predictive ability.At the same time,the calibration curve shows that the nomogram has a good degree of calibration in both the training set and the validation set.Decision curves found that the model was more beneficial than other factors in terms of net clinical benefit.Conclusions:(1).ACC of the breast cannot be simply summarized as triple-negative breast cancer because it also includes a small number of hormone receptor-positive breast cancers.(2).It is difficult to determine the preoperative diagnosis by clinical manifestations and imaging examinations,and FNAC may be a useful tool for diagnosis.(3).The proportion of breast-conserving patients in this hospital is lower than that in the SEER database.(4)The nomogram model constructed based on the independent prognostic factors of age,histological grade and N stage can effectively predict the overall survival rate of patients with breast adenoid cystic carcinoma,and has certain reference value for guiding treatment and evaluating the prognosis of patients. |
PDF Full Text Request