Atopic dermatitis is defined as an intensely systemic inflammation among skin diseases. Exosomes derived from adipose-derived stem cells may be a novel cell-free therapeutic strategy for atopic dermatitis treatment. This study aims to elucidate the possible underlying mechanism of adipose-derived stem cells-exosomes harboring microRNA-147a in atopic dermatitis pathogenesis. BALB/c mice treated with Dermatophagoides farinae extract/2,4-dinitrochlorobenzene were defined as a mouse model of atopic dermatitis, either with inflamed HaCaT cells and HUVECs exposed with TNF-α/IFN-γ stimulation were applied for a cell model of atopic dermatitis. The concentrations of IL-1β and TNF-α in the supernatants were examined by ELISA. Cell viability and migration were assessed by MTT and Transwell assay. The apoptosis was examined using flow cytometry and TUNEL staining. The tube formation assay was employed to analyzed angiogenesis. The molecular regulations among miR-147a, MEF2A, TSLP and VEGFA were confirmed using luciferase reporter assay, either with ChIP. microRNA-147a was markedly downregulated in the serum and skin samples of atopic dermatitis mice, of which overexpression remarkably promoted HaCaT cell proliferation, meanwhile inhibiting inflammatory response and cell apoptosis. microRNA-147a in adipose-derived stem cells was subsequently overexpressed, and exosomes (Exos-miR-147a mimics) were collected. Functionally, exos-microRNA-147a mimics attenuated TNF-α/IFN-γ-induced HaCaT cell inflammatory response and apoptosis, and suppressed HUVECs angiogenesis. Encouraging, molecular interaction experiments revealed that exosomal microRNA-147a suppressed TNF-α/IFN-γ-induced HUVECs angiogenesis by targeting VEGFA, and exosomal microRNA-147a repressed HaCaT cells inflammatory injury through the MEF2A-TSLP axis. Mechanistically, exosomal microRNA-147a repressed pathological angiogenesis and inflammatory injury during atopic dermatitis progression by targeting VEGFA and MEF2A-TSLP axis. microRNA-147a-overexpressing adipose-derived stem cells-derived exosomes suppressed pathological angiogenesis and inflammatory injury in atopic dermatitis by targeting VEGFA and MEF2A-TSLP axis.
In recent years, studies have found that miR-RNA plays a role in cell proliferation, differentiation, apoptosis and metabolism. Among them, miR-196a is closely related to cervical cancer. Therefore, this experiment investigates the effect of mir-196a expression on cervical cancer cells and related mechanisms. The expression level of miR-196a in the cervical cancer cell line was assayed with the RT-PCR method, and liposome transfection was used to investigate its up-regulation or down-regulation in cervical cancer cells. The CCK-8 method and flow cytometry were used to measure cervical cancer cell proliferation and apoptosis, while the Transwell assay was used to determine cell migration and invasion of each transfection group. Bioinformatics was used to predict the target gene of miR-196a, which was verified using dual luciferase report experiment and Western blot, and miR-196a was further transfected with si-LRIG3 to detect its reversal effect on miR-196a regulation. Inhibition of the expression of miR-196a significantly reduced the proliferation, migration and invasion of cervical cancer cells, and promoted their apoptosis. Results from dual luciferase assay showed that miR-196a and LRIG3 had direct targeting effects. Cell proliferation, migration and invasion were enhanced by a reduction in the expression level of LRIG3 protein after miR-196a inhibitor cells were transfected with si-LRIG3. The expression of miR-196a is up-regulated in cervical cancer, and it promotes the growth of cervical cancer by its targeting effect on LRIG3 expression, resulting in enhancement of the proliferation, migration and invasion ability of cervical cancer cells, and inhibition of apoptosis.
Myeloid leukemia-1 (Mcl-1) gene has been reported as an important factor in various types of cancer, but little research was processed on natural killer (NK)/T-cell lymphoma, a kind of a highly aggressive disease with a poor prognosis. Here we investigated the expression of Mcl-1 in seven lymphoma cell lines and its potential role as a molecular drug target for NK/T-cell lymphoma therapy by using lentivirus-mediated shRNA interference targeting Mcl-1 (lenti-shMcl-1). In our study, the expression of Mcl-1 in different lymphoma cell lines were evaluated firstly, after that lenti-shMcl-1 was constructed and transduced into NK/T-cell lymphoma cell line SNK-6 which had a high level expression of Mcl-1. Methylthiazolete-Trazolium (MTT) assay and flow cytometry (FCA) were employed to detect the status of proliferation and apoptosis after infection. Lastly we investigated the effects of chemotherapy agent vincristine (VCR) combination with lenti-shMcl-1 by MTT and FCA assay. The results showed that Mcl-1 gene expressed in all seven lymphoma cell lines at different levels. Recombinant lentiviruses could infect SNK-6 cells effectively. Mcl-1 expression level was remarkably down-regulated after infection with lenti-shMcl-1. The growth of SNK-6 cells was inhibited significantly through apoptosis pathway. Otherwise, lenti-shMcl-1 also revealed a significant chemosensitizing effect in combination with vincristine. In a word, we demonstrated that lenti-shMcl-1 had a significant anti-NK/T cell lymphoma effect and targeting Mcl-1 therapy could be a promising novel approach in treatment of lymphoma.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.