This study was undertaken to evaluate whether exosomes from human adipose-derived stem cells (ASC-exo) can stimulate the regeneration of human dermal fibroblasts (HDFs). Immunoblotting and FACS analyses showed that ASC-exo was positive for exosome markers. Fluorescence tracking revealed that the contents of ASCexo were transferred into the HDFs. ASC-exo treatment also stimulated the proliferation and migration of HDFs in a dose-dependent manner. Similarly, the expression levels of genes involved in skin cell proliferation were increased by ASC-exo. Microarray analysis showed an enrichment of microRNAs that have regenerative function. We suggest that the ASC-exo can stimulate skin cell proliferation. | BACKGROUNDIt has been well reported that adipose tissue-derived stem cell (ASC) secretes various soluble factors, that is growth factors, cytokines, exosomes that can rescue damaged cells.[1] Exosome, a 30-to 150-nm-sized extracellular vesicle that is secreted from most cell types, is formed within endosomal compartments and released into the extracellular milieu.[2] It has been demonstrated that exosomes possess similar functional properties of mesenchymal stem cells (MSCs) from which they are derived, [3] and due to such functions, exosomes are regarded now as a communicator between tissues.[4] Functionally, exosomes can reduce the immune recognition, so that the integrity of cell membrane can be maintained. [5] A study of stem cell transplantation therapy demonstrated that the role of MSCs in cell-tocell communication was exerted through a paracrine mechanism and that exosomes play a major role in this process.[6] Other findings also described that the conditioned media of ASC (ASC-CM) [7] or exosomes [8] were able to promote the migration of dermal fibroblasts during the process of wound healing. [7] Although the mechanism of the exosomes' role on proliferation and migration of fibroblasts was demonstrated, [8] no study has been conducted to identify the expressional changes of microRNAs, which play major role in various cellular responses, including proliferation. | QUESTIONS ADDRESSEDWe asked whether ASC-derived exosomes can stimulate the skin dermal fibroblasts' proliferation and migration, and by what mechanism these exosomes play such functions. | EXPERIMENTAL DESIGNSee supplementary information. | RESULTSTransmission electron microscopy (TEM) and nanoparticle tracking analysis (NTA) showed that ASC-exo isolated from the supernatants (Fig. S1A). Western blot analysis revealed that ASC-exo expresses exosome markers HSP70, CD63and CD9 (Fig. S1B), while these markers were absent in ASC lysate.Flow cytometry analysis also showed that ASC-exos were positive for CD9, CD63 and CD81 (Fig. S1C). We next asked whether the contents of ASC-exos could be transmitted to HDFs. After co-incubation, for 24 hours, of PKH26-labelled ASC-exos with HDFs, it was found that red fluorescence of PKH26 was localized within the cytoplasm of HDFs (Fig. S1D), showing that ASC-exo can be internalized intoHDFs. Also, quantitati...

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Bcl-w a pro-survival member of the Bcl-2 protein family, is expressed in a variety of cancer types, including gastric and colorectal adenocarcinomas, as well as glioblastoma multiforme (GBM), the most common and lethal brain tumor type. Previously, we demonstrated that Bcl-w is upregulated in gastric cancer cells, particularly those displaying infiltrative morphology. These reports propose that Bcl-w is strongly associated with aggressive characteristic, such as invasive or mesenchymal phenotype of GBM. However, there is no information from studies of the role of Bcl-w in GBM. In the current study, we showed that Bcl-w is upregulated in human glioblastoma multiforme (WHO grade IV) tissues, compared with normal and glioma (WHO grade III) tissues. Bcl-w promotes the mesenchymal traits of glioblastoma cells by inducing vimentin expression via activation of transcription factors, β-catenin, Twist1 and Snail in glioblastoma U251 cells. Moreover, Bcl-w induces invasiveness by promoting MMP-2 and FAK activation via the PI3K-p-Akt-p-GSK3β-β-catenin pathway. We further confirmed that Bcl-w has the capacity to induce invasiveness in several human cancer cell lines. In particular, Bcl-w-stimulated β-catenin is translocated into the nucleus as a transcription factor and promotes the expression of target genes, such as mesenchymal markers or MMPs, thereby increasing mesenchymal traits and invasiveness. Our findings collectively indicate that Bcl-w functions as a positive regulator of invasiveness by inducing mesenchymal changes and that trigger their aggressiveness of glioblastoma cells.

We already had reported that Bcl-w promotes invasion or migration in gastric cancer cells and glioblastoma multiforme (GBM) by activating matrix metalloproteinase-2 (MMP-2) via specificity protein 1 (Sp1) or β-cateinin, respectively. High expression of Bcl-w also has been reported in GBM which is the most common malignant brain tumor and exhibits aggressive and invasive behavior. These reports propose that Bcl-w-induced signaling is strongly associated with aggressive characteristic of GBM. We demonstrated that Sp1 protein or mRNA expression is induced by Bcl-w using Western blotting or RT-PCR, respectively, and markedly elevated in high-grade glioma specimens compared with low-grade glioma tissues using tissue array. However, relationship between Bcl-w-related signaling and aggressive characteristic of GBM is poorly characterized. This study suggested that Bcl-w-induced Sp1 activation promoted expression of glioma stem-like cell markers, such as Musashi, Nanog, Oct4 and sox-2, as well as neurosphere formation and invasiveness, using western blotting, neurosphere formation assay, or invasion assay, culminating in their aggressive behavior. Therefore, Bcl-w-induced Sp1 activation is proposed as a putative marker for aggressiveness of GBM.

Objective Preeclampsia is clinically unpredictable and associated with adverse outcomes. Pregnant women with suspected preeclampsia require intensive monitoring or hospitalization for elevated sFlt‐1 (soluble fms‐like tyrosine kinase‐1) to PlGF (placental growth factor) ratios before symptoms arise. We aimed to determine the sFlt‐1/PlGF ratio's usefulness in predicting adverse pregnancy outcomes in preeclampsia. Methods From January 2017 to February 2019, we measured the sFlt‐1/PlGF ratio in 73 singleton pregnant women suspected of preeclampsia and classified them into three groups: low‐risk (sFlt‐1/PlGF ratio < 38, n = 19), intermediate (38 ≤ ratio < 85, n = 9), and high‐risk (ratio ≥ 85, n = 32). Results Although the low‐ and high‐risk groups both experienced weight gain during pregnancy, their body mass index (BMI) differed after pregnancy (p = 0.004). The number of women who had been taking antihypertensive medications for chronic hypertension since early pregnancy was higher in the low‐risk group (31.6% vs. 22.2%, 6.7%). The gestational weeks at birth were lower in the high‐risk group compared to that of the low‐risk group (32.0 weeks vs. 35.79 weeks, p < 0.001). In the high‐risk group, the average neonatal weight was significantly lighter (p = 0.021), and the period of stay in the neonatal intensive care unit was longer than that in the low‐risk group (p = 0.003). Conclusion The sFlt‐1/PlGF ratio is a useful indicator of preeclampsia severity and can be utilized as a prognostic marker.