ABSTRACT
This study examined the expression and potential mechanism of microRNA (miRNA)-424-5p in nasopharyngeal carcinoma (NPC). NPC tissues were collected from 40 patients who were enrolled in the study, and skin samples were collected from 26 healthy subjects during plastic surgery as controls. We performed various in vitro assays using miR-424-5p to examine its function in primary NPC-1 cells. Bioinformatics was employed to analyze potential target genes and signaling pathways of miR-424-5p. We found that miR-424-5p expression in NPC tissues is downregulated and negatively correlated with lymph node metastasis and clinical staging. Expression of miR-424-5p in NPC cells was also downregulated, and transfection with miR-424-5p mimics inhibited proliferation, migration, and invasion of NPC-1 cells. Bioinformatics identified the AKT3 gene as a potential target of miR-424-5p and dual luciferase assays confirmed this finding. Upregulation of AKT3 expression rescued the inhibitory effect of miR-424-5p on the proliferation, migration, and invasion. Our results suggest that miR-424-5p inhibited the proliferation, migration, and invasion of NPC cells by decreasing AKT3 expression.
Subject(s)
Humans , Male , Female , Adult , Middle Aged , Aged , Gene Expression Regulation, Neoplastic , Nasopharyngeal Neoplasms/metabolism , MicroRNAs/metabolism , Proto-Oncogene Proteins c-akt/metabolism , Nasopharyngeal Carcinoma/metabolism , Signal Transduction , Cell Movement , Nasopharyngeal Neoplasms/genetics , Blotting, Western , MicroRNAs/genetics , Cell Line, Tumor , Cell Proliferation , Proto-Oncogene Proteins c-akt/genetics , Real-Time Polymerase Chain Reaction , Nasopharyngeal Carcinoma/genetics , Neoplasm InvasivenessABSTRACT
Abstract: Background: Myeloid leukemia cutis is the terminology used for cutaneous manifestations of myeloid leukemia. Objective: The purpose of this study was to study the clinical, histopathological and immunohistochemical features of myeloid leukemia cutis. Methods: This was a retrospective study of clinical and pathological features of 10 patients with myeloid leukemia cutis. Results: One patient developed skin lesions before the onset of leukemia, seven patients developed skin infiltration within 4-72 months after the onset of leukemia, and two patients developed skin lesions and systemic leukemia simultaneously. Of these patients, five presented with generalized papules or nodules, and five with localized masses. The biopsy of skin lesions showed a large number of tumor cells within the dermis and subcutaneous fat layer. Immunohistochemical analysis showed strong reactivity to myeloperoxidase (MPO), CD15, CD43 and CD45 (LCA) in most cases. NPM1 (nucleophosmin I) and FLT3-ITD (Fms-like tyrosine kinase 3-internal tandem duplication) mutations were identified in one case. Five patients with acute myelogenous leukemia and one patient with chronic myelomonocytic leukemia died within two months to one year after the onset of skin lesions. Study limitations: This was a retrospective and small sample study. Conclusions: In patients with myelogenous leukemia, skin infiltration usually occurs after, but occasionally before, the appearance of hemogram and myelogram abnormalities, and the presence of skin infiltration is often associated with a poor prognosis and short survival time. myeloid leukemia cutis often presents as generalized or localized nodules or masses with characteristic pathological and histochemical findings.
Subject(s)
Humans , Male , Female , Adult , Middle Aged , Aged , Aged, 80 and over , Skin/pathology , Leukemia, Myeloid/pathology , Leukemic Infiltration/pathology , Prognosis , Time Factors , Biopsy , Immunohistochemistry , Sex Factors , Retrospective Studies , Age FactorsABSTRACT
<p><b>OBJECTIVE</b>To evaluate the efficacy of mutation of human papillomavirus 16 (HPV16) E7 in two zinc-binding motifs on HPV16 E7 C terminus on antigen-specific immunity.</p><p><b>METHODS</b>pcDNA3.1/16E7 and pcDNA3.1/ME7 were successfully constructed by inserting the E7 (ME7) into pcDNA3.1 BamH I, EcoR I cut sites. After intramuscular injection with pcDNA3.1, pcDNA3.1/16E7, and pcDNA3.1/16ME7 on C57BL/6 mice, splenocytes from vaccinated mice was isolated. After have been stimulated with E7-specific peptide, interferon gamma (IFN-gamma), interleukin 2 (IL-2), and cytotoxic T lymphocyte (CTL) were detected by ELISA, Eli-spot, and LDH assay respectively; splenocytes without E7 peptide stimulation were used as control group.</p><p><b>RESULTS</b>Splenocytes from mice vaccinated with pcDNA/ME7, stimulated with E7 peptide, generated significantly larger number of E7-specific IL-2 compared with pcDNA3.1/16E7, pcDNA3.1, and control group. The E7-specific IL-2 generated in pcDNA-ME7 group was 5-fold of that of pcDNA3.1/16E7, and the difference was statistically significant (P < 0.05). Splenocytes from mice vaccinated with pcDNA/ME7 and stimulated with E7 peptide, generated significantly larger number of E7-specific IFN-gamma compared with other vaccines. pcDNA-ME7 generated a 2-fold increase in the number of E7-specific IFN-gamma compared with wild-type E7 and the difference was statistically significant (P < 0.05). The highest CTL activity in mice vaccinated with pcDNA/ME7 at an E:T ratio of 45:1 was achieved compared with mice vaccinated with other vaccines. The percents of specific lysis generated by pcDNA3.1/ME7, pcDNA3.1/E7, pcDNA3.1, and without vaccination were of (28.7+/-1.2) %, (55+/-2.2) %, (12.5+/-2.0) %, and (11.5+/-1.2) % respectively, and significant difference existed between the former and the latter two groups (P < 0.05). However, no significant difference was found among all the groups without specific E7 peptide stimulation (P > 0.05).</p><p><b>CONCLUSIONS</b>The mutation of zinc-binding motifs on HPV16 E7 C terminus may greatly enhance the immunogenicity.</p>