ABSTRACT
The most common mutations in gastrointestinal stromal tumors (GISTs) are KIT or PDGFRA mutations. Recently, neurotrophic tyrosine receptor kinase (NTRK) fusions have been reported in WT GISTs, which increased interest in introducing tropomyosin receptor kinase (TRK) inhibitors as treatments for GISTs with NTRK fusions. Hence, we aimed to screen NTRK fusions in WT GISTs; we used fluorescence in situ hybridization (FISH), next-generation sequencing (NGS), and immunohistochemistry (IHC) to screen NTRK fusions in 46 WT GISTs and evaluate each method. We further reviewed NTRK fusion-positive GISTs from the literature and performed clinical and pathological analyses; two GISTs with an ETV6-NTRK3 fusion (5%) were identified, while only one (50%) was positive for Pan-TRK expression. On the other hand, among the six GISTs with Pan-TRK-positive expression, only one (17%) harbored NTRK fusion. The literature review revealed the strong consistency between FISH and NGS and the limited value of Pan-TRK IHC in screening NTRK fusions in GISTs. In addition, the clinical and pathological analysis showed that GISTs with NTRK rearrangement occurred less frequently in the stomach, were more frequently larger in size, and the epithelioid type presented with a higher risk of recurrence. The NTRK3 fusion has been more common than the NTRK1 fusion in GISTs to date; our study identified two ETV6-NTRK3 fusions in 46 WT GISTs. Compared with FISH and IHC, NGS is preferred for screening WT GISTs, including NTRK rearrangements. However, since GISTs with NTRK fusions are rare, further studies including more samples and mechanistic investigations should be conducted in the future.
ABSTRACT
High expression of PD-L1 marks the poor prognosis of pancreatic ductal adenocarcinomas (PDAC). However, the regulatory mechanism of PD-L1 remains elusive. We recently reported that cancer Forkhead box protein 3 (Cancer-FOXP3 or C-FOXP3) promoted immune evasion of PDAC by recruiting Treg cells into PDAC via upregulation of CCL5. In this study, we confirmed that PD-L1 was overexpressed in PDAC samples from two independent cohorts of patients with radical resection. Moreover, C-FOXP3 was colocalized and correlated with the expression of PD-L1 in tumor cells at the mRNA and protein levels, and this finding was confirmed by the The Cancer Genome Atlas (TCGA) database. Chromatin immunoprecipitation (ChIP) revealed that C-FOXP3 directly bound to the promoter region of PD-L1 in pancreatic cancer cells. Furthermore, overexpression of C-FOXP3 activated the luciferase reporter gene under the control of the PD-L1 promoter. However, mutation of the binding motif-a completely reversed the luciferase activity. In addition, C-FOXP3-induced upregulation of PD-L1 effectively inhibited the activity of CD8+ T cells. Based on our recent finding that the CCL-5 antibody achieved a better response to PDAC models with high C-FOXP3 levels, we further demonstrated that the PD-L1 antibody strengthened the antitumor effect of CCL-5 blockade in xenograft and orthotopic mouse models with high C-FOXP3 levels. In conclusion, C-FOXP3 directly activates PD-L1 and represents a core transcription factor that mediates the immune escape of PDAC. Combined blockade of PD-L1 and CCL-5 may provide an effective therapy for patients with PDAC that have high C-FOXP3 levels.
Subject(s)
Adenocarcinoma/immunology , B7-H1 Antigen/genetics , Carcinoma, Pancreatic Ductal/immunology , Chemokine CCL5/genetics , Forkhead Transcription Factors/genetics , Adenocarcinoma/genetics , Adenocarcinoma/pathology , Adenocarcinoma/therapy , Aged , Animals , B7-H1 Antigen/antagonists & inhibitors , B7-H1 Antigen/immunology , CD8-Positive T-Lymphocytes/immunology , Carcinoma, Pancreatic Ductal/genetics , Carcinoma, Pancreatic Ductal/pathology , Carcinoma, Pancreatic Ductal/therapy , Chemokine CCL5/antagonists & inhibitors , Female , Gene Expression Regulation, Neoplastic/drug effects , Heterografts , Humans , Immune Checkpoint Inhibitors/pharmacology , Immunotherapy/methods , Lymphocytes, Tumor-Infiltrating/drug effects , Lymphocytes, Tumor-Infiltrating/immunology , Male , Mice , Middle Aged , Prognosis , T-Lymphocytes, Regulatory/immunologyABSTRACT
Most cancers are caused by somatic mutations. Some common mutations in the same cancer type can form a "signature" to specifically predict the prognosis or to distinguish it from other cancers. In this study, 710 somatic cell mutations were identified in 142 cases, including digestive, lung and urogenital cancers, and the digestive cancers were further divided into liver, stomach, intestinal, esophageal and cardia cancer. The above mutations were located in 166 genes. In addition, a group of high-frequency mutation genes with specific characteristics were screened to form predictive signatures for each cancer. Verification using TCGA suggested that the signatures could predict the stages, progression-free survival, and overall survival of digestive, intestinal, and liver cancers (P < 0.05). The validation cases further confirmed the predictive role of digestive and liver cancers signatures in diagnosis and prognosis. Overall, this study established predictive signatures for different cancer systems and their subtypes. These findings enable a better understanding in cancer genome, and contribute to the personalized diagnosis and treatment.
Subject(s)
Biomarkers, Tumor/genetics , DNA Mutational Analysis , Digestive System Neoplasms/diagnosis , Gene Expression Regulation, Neoplastic , Adult , Aged , Aged, 80 and over , Biomarkers, Tumor/analysis , Digestive System Neoplasms/genetics , Digestive System Neoplasms/mortality , Digestive System Neoplasms/therapy , Female , High-Throughput Nucleotide Sequencing , Humans , Male , Middle Aged , Mutation , Neoplasm Staging , Precision Medicine , Predictive Value of Tests , Prognosis , Progression-Free Survival , Reproducibility of Results , Young AdultABSTRACT
Small-cell lung cancer (SCLC) is an aggressive malignancy characterized by high cellular proliferation and early distant metastasis. Our study aimed to explore the effect of miR-22-3p (miR-22, for short) on SCLC radiosensitivity and its molecular mechanisms. The expression level of miR-22 was evaluated in a human normal lung epithelial cell line and a human SCLC cell line, and cell apoptosis and migration were detected. The expression of the miR-22 direct target WRNIP1 mRNA and protein were explored. Five differentially expressed genes were detected. The miR-22 expression in NCI-H446 was significantly decreased, and miR-22 overexpression significantly promoted cell apoptosis. miR-22 overexpression could significantly inhibit the cell migration of SCLC cells, and miR-22 had a negative regulatory effect on WRNIP1 mRNA and protein levels. KLK8 was downregulated, and the messenger RNA (mRNA) of four other genes (PC, SCUBE1, STC1, and GPM6A) was upregulated mRNA in cells overexpressing miR-22, which was in accordance with the bioinformatics analysis. miR-22 could enhance the radiosensitivity of SCLC by targeting WRNIP1.
Subject(s)
ATPases Associated with Diverse Cellular Activities/metabolism , DNA-Binding Proteins/metabolism , Lung Neoplasms/genetics , MicroRNAs/metabolism , Radiation Tolerance/genetics , Small Cell Lung Carcinoma/genetics , 3' Untranslated Regions/genetics , ATPases Associated with Diverse Cellular Activities/genetics , Apoptosis/genetics , Base Sequence , Cell Line, Tumor , Cell Movement/genetics , Cell Proliferation/genetics , DNA-Binding Proteins/genetics , Dose-Response Relationship, Radiation , Gene Expression Regulation, Neoplastic , Gene Knockdown Techniques , HEK293 Cells , Humans , Ki-67 Antigen/metabolism , Lung Neoplasms/pathology , MicroRNAs/genetics , Small Cell Lung Carcinoma/pathologyABSTRACT
Osteoarthritis (OA) is a common type of disease affecting the joints that results from the breakdown of joint cartilage and the underlying bone; currently, its pathogenesis is still unclear. The aim of the present study was to identify key mRNAs and miRNAs involved in the pathogenesis and progression of OA using microarray analysis. The gene expression profile of GSE27492 was downloaded from the Gene Expressed Omnibus database, and included 49 arthritic mouse ankle samples collected at 6 time points (0, 1, 3, 7, 12 and 18 days) following the induction of arthritis via serum transfer. Differentially expressed genes (DEGs) were identified in ankle samples taken on days 1, 3, 7, 12 and 18 following serum transfer compared with day 0 samples, and overlapping DEGs in day 3, 7, 12 and 18 samples were identified. The Database for Annotation, Visualization and Integrated Discovery online tool was used to perform functional and pathway enrichment analyses of the overlapping DEGs. The miRWalk database was used to identify potential micro (mi) RNAs regulating the selected overlapping DEGs, and regulatory miRNAtarget mRNA pairs were obtained. The Cytoscape platform was used to establish and visualize the miRNAmRNA regulatory network. The present results revealed that 35, 103, 62 and 75 DEGs were identified in day 3, 7, 12 and 18 samples, respectively. A total of 17 overlapping DEGs were identified among the 4 sample sets, and revealed to be enriched in 14 gene ontology terms and 3 Kyoto Encyclopedia of Genes and Genomes pathways. miRWalk analysis identified 242 potential miRNAmRNA regulatory pairs and 211 nodes were revealed to be involved in the miRNAmRNA regulatory network. The present study identified potential genes, including Ctype lectin domain family 4 member D, chemokine (CXC motif) ligand 1 and CC motif chemokine ligand, and pathways, including chemokine signaling pathways, cytokinecytokine receptor interactions and nucleotidebinding oligomerization domainlike receptor signaling pathways, which may be involved in the pathogenesis and progression of OA. These findings may help elucidate the molecular mechanisms underlying OA pathophysiology, and may be useful for the development of novel therapeutic targets for the treatment of patients with OA.
Subject(s)
Gene Expression Regulation , MicroRNAs/genetics , Osteoarthritis/genetics , Osteoarthritis/pathology , RNA Interference , RNA, Messenger/genetics , Computational Biology , Databases, Genetic , Disease Progression , Gene Expression Profiling , Gene Regulatory Networks , Humans , Osteoarthritis/metabolism , Signal TransductionABSTRACT
OBJECTIVE: To study transfection efficiency of Ad5F11p-GFP and its influence on biological characteristics of CIK and NK-92 cells in order to predict the application of Ad5F11p vector in immunotherapy. METHODS: Two kinds of immune cells, cytokine-induced killer (CIK) cells and natural-killer (NK) cell line NK-92 cells, were transfected by Ad5F11p-GFP at different multiplicity of transfection (MOI), and untransfected immune cells were used as negative control. GFP expression was determined by flow cytometry, the cell morphology was observed with microscope, the cell proliferation was analyzed by trypan blue staining, specific cytotoxicity of NK-92 cells was determined by LDH assay. RESULTS: About 90% of transfection efficiency for NK-92 cells could be achieved at a MOI of 25, while the transfection efficiency for CIK was less than 40% at a MOI of 200. In addition, the transfection efficiency basically unchanged at the same MOI for 48 h and 96 h, and the immune cells transfected with the virus trended to form agglomeration, displaying slower proliferation, increase of IFN-γ release and enhancement of tumor killing activity. CONCLUSION: Ad5F11p- modified NK-92 shows a good prospect for adoptive immunotherapy.