ABSTRACT
Objective:To investigate changes of nicastrin (NCSTN) downstream molecules in signaling pathways related to cell proliferation and differentiation after silencing the expression of the NCSTN gene in the human immortalized keratinocyte cell line HaCaT.Methods:HaCaT cells were divided into 3 groups: interference group transfected with a specific small interfering RNA (siRNA) targeting NCSTN (NCSTN-siRNA) , negative control group transfected with a negative control siRNA, and blank control group transfected with the equal amount of transfection reagent. Real-time PCR and Western blot analysis were performed to measure the NCSTN mRNA and protein expression in groups, in order to verify the transfection efficiency. Differences in gene expression profiles in HaCaT cells were detected between the interference group and negative control group by using Agilent whole-genome microarray, and differentially expressed genes were identified based on a fold change ≥ 2.0 with a P value ≤ 0.05. Gene ontology (GO) enrichment analysis was employed to identify the roles of the differentially expressed genes, and then to screen out significantly differentially expressed genes associated with proliferation and differentiation of keratinocytes, some of which were verified by real-time PCR. Results:The interference group showed significantly decreased mRNA and protein expression of NCSTN (0.287 ± 0.090, 0.443 ± 0.085, respectively) compared with the negative control group (0.969 ± 0.127, 1.047 ± 0.114, respectively) and blank control group (1.000 ± 0.151, 1.000 ± 0.111, F = 30.787, 31.139, respectively, both P = 0.001) . Whole genome-expression analysis using an Agilent microarray platform revealed 605 downregulated genes and 444 upregulated genes in HaCaT cells in the interference group compared with the negative control group. GO analysis showed that differentially expressed genes were enriched into 4 biological processes, including epithelial development, epithelial cell differentiation, keratinocyte differentiation and keratinization. The significantly differentially expressed genes associated with proliferation and differentiation of keratinocytes, including the Sprouty-related protein with EVH1 domain 2, fibroblast growth factor 7, insulin-like growth factor-binding protein 5, Rho-associated coiled-coil kinase 2 and bone morphogenetic protein 6 genes, were verified by real-time PCR, and the verification results were consistent with the difference trend shown by the microarray results. Conclusion:The loss of NCSTN gene function may affect the normal proliferation and differentiation of keratinocytes by regulating the expression of its downstream molecules in signaling pathways associated with cell proliferation and differentiation.
ABSTRACT
Acne inversa is a chronic inflammatory skin disease of the folliculo-sebaceous-apocrine unit. Currently, genetic and immunological factors are hot topics in the study of its pathogenesis. Genetic factors are mainly related to γ-secretase mutations, and abnormal expression of the γ-secretase-Notch axis leads to increased keratinization of hair follicles and inflammation in some patients with haploinsufficiency of the γ-secretase gene. Mutations in the γ-secretase gene are not necessary for acne inversa, and the risk of Alzheimer′s disease in familial acne inversa patients still remains unclear. Some progress has been made in researches on the association of genotype with phenotype in acne inversa patients, but further studies with large sample size are needed for verification.
ABSTRACT
Acne inversa is a chronic inflammatory skin disease of the folliculo-sebaceous-apocrine unit.Currently,genetic and immunological factors are hot topics in the study of its pathogenesis.Genetic factors are mainly related to γ-secretase mutations,and abnormal expression of the γ-secretase-Notch axis leads to increased keratinization of hair follicles and inflammation in some patients with haploinsufficiency of the y-secretase gene.Mutations in the γ-secretase gene are not necessary for acne inversa,and the risk of Alzheimer's disease in familial acne inversa patients still remains unclear.Some progress has been made in researches on the association of genotype with phenotype in acne inversa patients,but further studies with large sample size are needed for verification.
ABSTRACT
A 1-year-old boy developed multiple skin-colored nodules on the forehead and extremities when he was 4 months old.Physical examination revealed that his general condition was well with no hepatomegaly,splenomegaly,lymphadenectasis,testicle abnormality or gingival hypertrophy.Pathologically,the epidermis was normal,while the dermis and subcutaneous tissue were diffusely infiltrated with medium-to large-sized deformed cells,which had a small amount of cytoplasm,oval nucleus,irregular shape and fine chromatin.Some infiltrating cells had nuclear groove and nucleoli.Immunohistochemical studies showed that the tumor cells were positive for S-100 protein,CD56,CD123,CD163,CD68,Ki-67 (40％),weakly positive for CD4 (some),but negative for myeloperoxidase,CD1,CD21.Bone marrow smears showed a 24.5％ infltration by monoblasts and promonocytes.A diagnosis of monoblastic sarcoma cutis preceding acute monoblastic leukemia was made.
ABSTRACT
Objective To study the role of serum chemokines and their receptors in the pathogenesis of atopic dermatitis(AD).Methods Serum levels of interferon ?-inducible protein-10(IP-10),stromal cell-derived factor-1? (SDF-1?),eotaxin,thymus and activation-regulated chemokine(TARC) as well as macrophage-derived chemokine (MDC) were detected by enzyme-linked immunosor bent assay(ELISA) in 39 patients with AD and 39 normal controls.Meanwhile,chem okine receptors CXCR3,CXCR4,CCR3,CCR4 and CCR5 on peripheral blood CD4+T-lymp hocytes were analyzed by flow cytometry with a two-color immunofluorescent sta ining in 39 patients with AD.Results Serum levels of SDF-1?,TARC and MDC we re significantly higher(P 0.05) between the patients[(123.6 ? 110.4) pg/mL a nd (68.7 ? 26.2) pg/mL,respectively]and controls [(100.7 ? 73.7) pg/mL and(66.8 ? 20.5) pg/mL,respectively].The expression of CXCR3,CCR3,CCR4 and CCR5 o n CD4+T cells was increased (P