| Objective(s):(1)The clinicopathological and follow-up data of patients with aggressive fibromatosis(AF)were collected from multiple centers to establish a tumor paraffin-embedded sample bank of AF patients,and to analyze the key factors related to the prognosis of AF patients.(2)Proteomics was used to detect the complete protein expression profile in the sample bank,compare the protein expression differences in different subgroups of AF patients,and find the key proteins related to prognosis.Bioinformatics information was used to analyze the relationship between key proteins and the occurrence and development of AF tumors,so as to reveal the pathogenesis of this tumor and provide theoretical basis for the development of new therapeutic drugs.Methods:1.From January 2016 to December 2021,AF patients who underwent surgery and were diagnosed by pathologists were enrolled in this multicenter study.The relevant clinicopathological data were collected.Color Doppler ultrasound or MRI was reviewed at least once a year to determine tumor recurrence,and MRI was reviewed at least once every 6 months to observe tumor growth after recurrence.The patients were divided into non-recurrence group(NR group),stable recurrence group(RS group),increased recurrence group(RE group)and reduced recurrence group(RR group).SPSS 24.0 software was used to describe the data.log-rank single factor and Cox regression multivariate analysis were used to analyze the key factors affecting the recurrence-free survival of AF patients.2.The postoperative paraffin sections of AF patients were collected for proteomics detection by liquid chromatography tandem mass spectrometry(LC-MS/MS).data independent acquisition(DIA)was used for data analysis.Firstly,after the samples were reduced,alkylated and digested by Trypsin,the sample concentration was detected for correction and quality control analysis.Then,the database was established by traditional data dependent acquisition(DDA)technology,and the samples were analyzed by DIA model.The above DDA database was used for qualitative and quantitative analysis.Related bioinformatics analysis included:(1)quantitative analysis of identified proteins: total protein quantity and quantitative analysis,PCA analysis of samples,venn analysis of samples between groups;(2)Analysis of differentially expressed proteins: quantitative analysis,volcano plot and cluster analysis;(3)Biological function analysis of related proteins: subcellular localization(SL)analysis,domain analysis,GO function and KEGG pathway analysis,protein interaction network(PPI)analysis.Results:(1)97 patients with AF were included in the study.The follow-up time was from July to 86 months,with a median follow-up time of 24 months.There were 29 males and 68 females;The median age of onset was 29 years;32 patients had lesions located in the extremities,14 in the abdominal cavity,15 in the abdominal wall,and 36 in the trunk.26 patients with tumor diameter < 5cm,46 patients with tumor diameter < 5cm and 46 patients with tumor diameter < 10 cm,25 patients with tumor diameter ≥10cm;37 patients had vascular nerve involvement and 60 patients had non-vascular nerve involvement.Ki67 protein expression < 5% in 61 patients and ≥5% in 12 patients;β-catenin protein was expressed in 82 patients and not expressed in 3 patients;47patients recurred after surgery,13 patients recurred multiple times;The 2-year RFS of tumor were 61.6% and the 5-year RFS were 40.2%.In log-rank univariate analysis,primary tumor size(P < 0.001),vascular or nerve involvement(P = 0.005),ki-67(P =0.007)were correlated with the recurrence of AF tumors.Cox regression multifactorial analysis showed that tumor size was an independent factor affecting RFS in AF patients(P = 0.019).(2)571 significantly different proteins were identified between the RS,RE,andRE groups and the NR group.The differential proteins were mainly concentrated in the down-regulated proteins,with 37 down-regulated proteins and 1 up-regulated protein,which were mainly located in the nucleus.The first three biological processes in which differential proteins were significantly enriched by GO analysis were monosaccharide metabolism,bacterial defense and antibacterial body fluids,etc.The first three categories of molecular functions were the structural components of muscle cells,the activity of structural molecules and cytoskeleton-binding proteins.The first three categories significantly enriched in cellular components were immunoglobulin IgA complex,contractile fiber,and myofibrillar fiber.KEGG pathway analysis showed that the different proteins were mainly concentrated in glycolysis/glucose generation pathway,glucagon signaling pathway,AMPK signaling pathway and tumor micro RNA regulatory pathway.The protein interaction network analysis was performed for 38 different proteins,combined with the protein interaction score,and ROC characteristic curve analysis was performed for all the different proteins.It was found that the expression level of ARMT1 protein had predictive value for AF recurrence.Kaplan-Meier survival curves of patients with different expression levels of ARMT1 protein were compared,and the 5-year RFS of patients with high expression of ARMT1 protein were 50% and those with low expression of ARMT1 protein were 26%,showing a statistical difference(P=0.005).(3)Stability(RS group),enlargement(RE group),and shrinkage(RR group)of AF tumor after recurrence were also compared between groups.In terms of the number of significantly different proteins,12 up-regulated proteins and 31down-regulated proteins were identified between RS and RE groups.590 up-regulated proteins and 17 down-regulated proteins were identified between RR and RE groups.12 up-regulated proteins and 683 down-regulated proteins were identified between RS and RR groups.The analysis of differential proteins between RS and RE groups showed that the proteins with higher upregulation ratios were prickly protein 4 and interferon-related developmental regulatory factor 2And so on.The analysis of differential proteins between RR and RE groups showed that the proteins with high upregulation ratio,such as metalloproteinase 3,GAP repeat protein 1,and immunoglobulin 2-11,could inhibit tumor growth through immunoregulation.The analysis of differential proteins between RS and RR showed that the down-regulated proteins with high differential ratios were mainly concentrated in the myosin family,such as myosin 1 to myosin 7,and related proteins,such as sarcoplasmic/endoplasmic reticulum calcium ATpase 2,ubiquitination-related proteins E3,ubiquitination-related proteins,etc.Subcellular localization analysis showed that the differential proteins between RS and RE groups were mainly concentrated in the nucleus and cell membrane,the differential proteins between RR and RE groups were mainly concentrated in the nucleus and cytoplasm,and the differential proteins between RS and RR groups were mainly concentrated in the nucleus and cytoplasm.GO analysis combined with KEGG pathway analysis showed that the differential proteins of RS and RE groups were mainly related to immune regulation function,the differential proteins of RR and RE groups were mainly related to metabolic regulation and immune regulation,and the differential proteins of RS and RR groups were mainly related to energy metabolism and metabolic abnormalities related to muscle protein.Conclusion(s):(1)Tumor size,vascular or nerve involvement,ki-67 were the related factors affecting tumor recurrence in patients with AF;Primary tumor size was an independent factor affecting RFS in patients with invasive fibroma.(2)ARMT1 protein is expected to be a molecular marker guiding the prognosis of AF.(3)The growth of relapsed tumor may be related to metabolic regulation. |
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