Humans ; Mutation ; Sarcoma/genetics* ; Ribonuclease III/genetics* ; DEAD-box RNA Helicases/genetics*

A complete proteomics study characterizing active androgen receptor (AR) complexes in prostate cancer (PCa) cells identified a diversity of protein interactors with tumorigenic annotations, including known RNA splicing factors. Thus, we chose to further investigate the functional role of AR-mediated alternative RNA splicing in PCa disease progression. We selected two AR-interacting RNA splicing factors, Src associated in mitosis of 68 kDa (SAM68) and DEAD (Asp-Glu-Ala-Asp) box helicase 5 (DDX5) to examine their associative roles in AR-dependent alternative RNA splicing. To assess the true physiological role of AR in alternative RNA splicing, we assessed splicing profiles of LNCaP PCa cells using exon microarrays and correlated the results to PCa clinical datasets. As a result, we were able to highlight alternative splicing events of clinical significance. Initial use of exon-mini gene cassettes illustrated hormone-dependent AR-mediated exon-inclusion splicing events with SAM68 or exon-exclusion splicing events with DDX5 overexpression. The physiological significance in PCa was investigated through the application of clinical exon array analysis, where we identified exon-gene sets that were able to delineate aggressive disease progression profiles and predict patient disease-free outcomes independently of pathological clinical criteria. Using a clinical dataset with patients categorized as prostate cancer-specific death (PCSD), these exon gene sets further identified a select group of patients with extremely poor disease-free outcomes. Overall, these results strongly suggest a nonclassical role of AR in mediating robust alternative RNA splicing in PCa. Moreover, AR-mediated alternative spicing contributes to aggressive PCa progression, where we identified a new subtype of lethal PCa defined by AR-dependent alternative splicing.
Humans ; Male ; Alternative Splicing ; Cell Line, Tumor ; DEAD-box RNA Helicases/metabolism* ; Disease Progression ; Gene Expression Regulation, Neoplastic ; Prostatic Neoplasms/pathology* ; Receptors, Androgen/metabolism* ; RNA Splicing Factors/metabolism*

OBJECTIVE: To analyze the clinical characteristics and genetic variant of a child featuring X-linked mental retardation. METHODS: Whole exome sequencing and Sanger sequencing were used for the detection of variant and pedigree validation, respectively. Clinical manifestation of patients with DDX3X gene variants were also reviewed. RESULTS: The child was found to harbor a heterozygous NM_001193416.3: c.1332_1333delCT (p.Leu445Serfs*19) variant of the DDX3X gene. The same variant was not found in either of her parents. CONCLUSION The child was diagnosed with X-linked mental retardation due to variant of the DDX3X gene. Above finding has enriched the spectrum of DDX3X gene variants and provided a basis for clinical diagnosis and prenatal diagnosis for this pedigrees.
Child ; DEAD-box RNA Helicases/genetics* ; Female ; Heterozygote ; Humans ; Intellectual Disability/genetics* ; Mental Retardation, X-Linked/genetics* ; Mutation ; Pedigree ; Pregnancy ; Exome Sequencing

OBJECTIVE: To develop a multiplex PCR method for a rapid detection of Y chromosome-specific sequences in patients with Turner syndrome. METHODS: Nine genes were selected from various regions of the Y chromosome for designing the primers, which included SRY, TBL1Y, TSPY on the short arm of the Y chromosome, DDX3Y, HSFY1, RPS4Y2 and CDY1 on the long arm of Y chromosome and SHOX in the short arm and SPRY3 in the long arm of the pseudoautosomal region (PAR) of X and Y chromosomes. A multiplex PCR method for the nine genes in Y chromosome was established and optimized. The sensitivity was tested by using different amounts of genomic DNA. A total of 36 patients with Turner syndrome and a patient with male dwarfism with karyotype of 46, X, +mar were examined by the multiplex PCR method for the existence of materials from the Y chromosome. RESULTS: The optimization results of the multiplex PCR reaction system (50 μL) showed that when the final concentration of upstream and downstream of each pair of primers was 0.1 μM, the multiplex PCR reaction of the 9 pairs of primers clearly amplified the target with the expected band size, and there was no non-specific amplification. The bands were clearly visible when the amount of genomic DNA in the multiple PCR reaction system was as low as 1 ng. By using the method, we have examined the 36 patients with Turner syndrome. One patient with Turner syndrome with karyotype of 45,X[40]/47XYY[21] amplified specific seven genes on Y chromosome, 35 patients with Turner syndrome amplified only two target genes SHOX and SPRY3, but not the other seven specific genes on the Y chromosome, which was in keeping with the clinical manifestations of such patients. CONCLUSION This study established a multiplex PCR reaction system with nine genes, which can quickly and accurately screen Y chromosome materials in patients with Turner syndrome. It has the advantages of low cost, simple operation, high specificity and rapid turn-around time, and can be used to detect Turner syndrome patients with Y chromosome material in time. The method has provided a diagnostic basis for preventive gonad resection to prevent malignant gonadal tumors.
Humans ; Male ; Turner Syndrome/genetics* ; Multiplex Polymerase Chain Reaction ; Y Chromosome ; Karyotyping ; DNA Primers ; DNA ; Chromosomes, Human, Y/genetics* ; Transducin/genetics* ; Minor Histocompatibility Antigens ; DEAD-box RNA Helicases/genetics*

Myelodysplastic syndrome (MDS) are a heterogeneous group of hematological malignancies. Currently, in addition to demethylated chemotherapy and hematopoietic stem cell transplantation, MDS patient-derived mesenchymal stem cells (MDS-MSC) play an important role in understanding the pathogenesis of MDS and related therapeutic targets. For example, abnormal expression of DICER1 gene, abnormalities of PI3K/AKT and Wnt/β-catenin signaling pathways provide new therapeutic targets for MDS. In addition, MDS-MSC is also affected by abnormal microenvironment of the body, such as inflammatory factor S100A9, as well as hypercoagulation and iron overload. In this review, genes, signaling pathways, cytokines, hematopoietic microenvironment, and the effect of therapeutic drugs for MDS-MSC were briefly summarized.
Cytokines/metabolism* ; DEAD-box RNA Helicases/metabolism* ; Hematologic Neoplasms/metabolism* ; Humans ; Mesenchymal Stem Cells ; Myelodysplastic Syndromes/genetics* ; Phosphatidylinositol 3-Kinases/metabolism* ; Ribonuclease III/metabolism* ; Tumor Microenvironment

OBJECTIVE: To explore the effective components of Yiqi Jiedu recipe and the main biological processes and signal pathways involved in the therapeutic mechanism of the recipe in treatment of primary liver cancer through network pharmacology and molecular docking approaches. METHODS: TCMSP, Uniport, Genecards and String databases were searched to obtain the target genes of drugs and disease using Cytoscape 3.8.2 software. GO and KEGG enrichment analyses were performed to identify the common genes in the target genes of the drugs and disease. Using Pubcham, RCSB and Autoduck, the effective components of the drugs were connected with the final core genes. The effects of different concentrations of Yiqi Jiedu recipe on the expressions of the core genes DHX9, HNRNPK, NCL and PABPC1 in HepG2 cells were analyzed with Western blotting and real- time fluorescence quantitative PCR. RESULTS: We finally identified 8 core genes from the drug and disease targets, including DDX5, HNRNPK, PABPC1, DHX9, RPS3A, RPS3, RPL13, and NCL. GO analysis showed that these core genes were involved mainly in the biological processes of adrenaline receptor signal communication, movement of cellular or subcellular components, blood particles, adhesion class and iron ion binding. KEGG analysis showed that the Ras signaling pathway had the greatest gene enrichment. The results of molecular docking suggested that the effective components of the recipe were capable of docking with the core genes under natural conditions, and PABPC1 and stigmasterol had the highest binding energy. In HepG2 cells, treatment with 10% medicated serum for 48 h had the strongest effect on the expression of DHX9, HNRNPK, NCL and PABPC1 (P < 0.05). CONCLUSION Yiqi Jiedu recipe is capable of regulating viral expression of primary liver cancer multiple effective components that bind to DHX9, HNRNPK, NCL and PABPC1.
DEAD-box RNA Helicases ; Drugs, Chinese Herbal/therapeutic use* ; Humans ; Liver Neoplasms/drug therapy* ; Molecular Docking Simulation ; Neoplasm Proteins ; Network Pharmacology ; Ribosomal Proteins ; Signal Transduction

OBJECTIVE: To analyze the clinical characteristics and treatment effects of children with acute megakaryoblastic leukemia without down syndrome (non-DS-AMKL). METHODS: The clinical data of 19 children with non-DS-AMKL treated in the Pediatric Hematology Ward in Sun Yat-sen Memorial Hospital of Sun Yat-sen University from May 2008 to April 2018 were analyzed retrospectively. The clinical characteristics, laboratory test and treatment methods of the children were concluded. All patients were followed up to evaluate the effect of treatment. RESULTS: The 19 cases of children included nine male and ten female, the median age of onset was 2 years old. The clinical manifestations showed nonspecific. The median white blood cell of peripheral blood was 15.88×10 CONCLUSION Non-DS-AMKL was rare in children and difficult to be diagnosed. Determination of MICM classification as early as possible was helpful for diagnosis, and genetic testing played an important role for diagnosis and prognosis evaluation. Early hematopoietic stem cell transplantation in patients with CR after chemotherapy might be an effective way to cure AMKL.
Child ; Child, Preschool ; DEAD-box RNA Helicases ; DNA Helicases ; Down Syndrome ; Female ; Humans ; Leukemia, Megakaryoblastic, Acute/genetics* ; Male ; Prognosis ; Retrospective Studies ; Trisomy

RNA helicases, the largest family of proteins that participate in RNA metabolism, stabilize the intracellular environment through various processes, such as translation and pre-RNA splicing. These proteins are also involved in some diseases, such as cancers and viral diseases. Autophagy, a self-digestive and cytoprotective trafficking process in which superfluous organelles and cellular garbage are degraded to stabilize the internal environment or maintain basic cellular survival, is associated with human diseases. Interestingly, similar to autophagy, RNA helicases play important roles in maintaining cellular homeostasis and are related to many types of diseases. According to recent studies, RNA helicases are closely related to autophagy, participate in regulating autophagy, or serve as a bridge between autophagy and other cellular activities that widely regulate some pathophysiological processes or the development and progression of diseases. Here, we summarize the most recent studies to understand how RNA helicases function as regulatory proteins and determine their association with autophagy in various diseases.
Animals ; Antiviral Agents/pharmacology* ; Autophagy ; Beclin-1/metabolism* ; Carcinogenesis ; Cell Survival ; DEAD Box Protein 58/metabolism* ; Disease Progression ; Gene Expression Regulation ; Homeostasis ; Humans ; Immune System/physiology* ; Neoplasms/metabolism* ; RNA Helicases/metabolism* ; RNA Splicing ; Receptors, Immunologic/metabolism*

OBJECTIVETo investigate the relationship between the expression of DDX39 protein and prognosis in colorectal cancer.

METHODSClinical data and paraffin specimens of postoperative tumor tissue from 824 patients with primary colorectal cancer who received first surgical treatment at the Department of Colorectal Surgery of Changhai Hospital of Navy Military Medical University from January 2010 to December 2011 were collected. Paraffin samples of paracancerous tissues of 38 patients were served as controls. At the same time, samples of normal rectal mucous membrane from 37 cases after procedure of prolapse and hemorrhoids, and samples of colorectal adenoma from 33 cases after endoscopic treatment were enrolled in this study. All the specimens were made as the tissue microarray, and the expression of DDX39 protein was detected by immunohistochemistry. The expression of DDX39 in the epithelium and stroma was evaluated with the average staining intensity (H-Score) and the number of positive cells. It was defined as high expression in the epithelium that the H-Score was greater than or equal to 200. It was defined as high expression in the stroma that the number of positive cells was greater than or equal to 50 in 200 times the field of vision. Relationship of different DDX39 expression levels with clinicopathological parameters and prognosis of colorectal cancer was analyzed.

RESULTSThe expression of DDX39 in colorectal cancer tissues was lower than that in normal tissues, paracancerous tissues and adenomatous tissues, whether it is in the epithelium or in the stroma [DDX39 expression in the epithelium: normal tissues 253.2±64.1, paracancerous tissues 238.8±79.2, adenomatous tissues 259.4±51.6, colorectal cancer tissues 194.2±76.5 (P=0.000, P=0.005, P=0.000, respectively); DDX39 expression in the stroma: normal tissues 110.1±64.8, paracancerous tissues 106.0±49.2, adenomatous tissues 108.5±79.1, colorectal cancer tissues 54.1±34.7(all P=0.000)]. Among the cases of colorectal cancer, there were 541 cases of high DDX39 expression and 283 cases of low DDX39 expression in the epithelium; there were 424 cases of high DDX39 expression of and 400 cases of low DDX39 expression in the stroma. The high DDX39 expression and low DDX39 expression in epithelial and stromal of colorectal cancer were related respectively with tumor location (P=0.006, P=0.016), degree of tumor differentiation (P=0.002, P=0.064), TNM stage (P=0.021, P=0.000), serum CEA level (P=0.003, P=0.005), serum CA199 level (P=0.040, P=0.005) and tumor recurrence and metastasis (P=0.000, P=0.000). All the colorectal cancer cases were followed up for (41.6±15.7) months after operation. The 5-year overall survival (OS) and disease-free survival (DFS) rates of the cases with epithelial low DDX39 expression were 84.1% and 61.5%, and both were significantly lower as compared to those with epithelial high DDX39 expression (95.4% and 88.2%, P=0.000, P=0.000). The 5-year OS and DFS rates of the stroma low DDX39 expression were 86.8% and 66.8%, and both were significantly lower as compared to those with stroma high DDX39 expression (96.1% and 90.6%, P=0.000, P=0.000). Cox multivariate analysis showed that tumor differentiation (OS:HR=0.252, 95%CI: 0.128 to 0.497, P=0.000; DFS:HR=0.266, 95%CI: 0.134 to 0.530, P=0.000), DDX39 expression level in epithelium (OS: HR =0.229, 95%CI: 0.138 to 0.382, P=0.000; DFS: HR =0.266, 95%CI: 0.158 to 0.446, P=0.000), and DDX39 expression level in stroma (OS: HR =0.331, 95%CI: 0.188 to 0.582, P=0.000; DFS:HR=0.326, 95%CI: 0.184 to 0.578, P=0.000) were independent influencing factors of overall or disease-free survival in patients with colorectal cancer.

CONCLUSIONThe low expression of DDX39 protein suggests poor prognosis and DDX39 is expected to be a new prognostic marker of colorectal cancer.

Biomarkers, Tumor ; metabolism ; Colonic Neoplasms ; Colorectal Neoplasms ; metabolism ; pathology ; DEAD-box RNA Helicases ; metabolism ; Disease-Free Survival ; Humans ; Neoplasm Recurrence, Local ; Neoplasm Staging ; Prognosis

Objective To investigate the effects of microRNA-18a (miR-18a) on migration and invasion of hepatocellular carcinoma (HCC) cells, and its possible mechanism associated with Dicer l.Methods HepG2 and HepG2.2.15 cells were transfected with miR-18a inhibitor using Lipofectamine. Cell invasion was evaluated by transwell invasion assay, and cell migration was detected by transwell migration and wound-healing assays. Moreover, luciferase reporter assay was used to identify whether Dicer expression was regulated by miR-18a. Real-time RT-PCR and western blot were performed to analyze Dicer 1 expression. In addition, a functional restoration assay was performed to investigate whether miR-18a promotes HCC cell migration and invasion by directly targeting Dicer 1.Results miR-18a inhibitor can suppress the migration and invasion of HCC cells. Furthermore, suppression of Dicer l expression by small interfering RNA essentially abolished the inhibition of cell migration and invasion induced by miR-18a inhibitor, restorating these activities to levels similar to the parental HCC cells. Interestingly, suppression of miR-18a in HCC cells resulted in enhanced expression of Dicer l. In addition, the results of a luciferase assay demonstrated targeted regulation of Dicer l by miR-18a.Conclusion Our findings suggest that miR-18a promotes migration and invasion of HCC cells by inhibiting Dicer l expression.
Carcinoma, Hepatocellular ; genetics ; metabolism ; pathology ; Cell Movement ; DEAD-box RNA Helicases ; genetics ; metabolism ; Hep G2 Cells ; Humans ; Liver Neoplasms ; genetics ; metabolism ; pathology ; MicroRNAs ; genetics ; metabolism ; Neoplasm Invasiveness ; Neoplasm Proteins ; genetics ; metabolism ; RNA, Neoplasm ; genetics ; metabolism ; Ribonuclease III ; genetics ; metabolism