Selenium Dioxide Induced Apoptosis in Cervical Cancer Cells via Regulating Apoptosis-related Let-7a MicroRNA and Proteins

Aim: To investigate the effects of selenium dioxide (SeO2) on the apoptosis and the involved epigenetic mechanisms in two cervical cancer cell lines. Methods: Human cervical carcinoma cell lines HeLa and CaSki were treated with 1.875–30 µmol/L SeO2 for 24 h. Morphological changes were observed by optical microscope; anti-proliferative effects were examined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay; flow cytometry (FCM) was employed to detect the cell apoptosis; real-time polymerase chain reaction was used to detect the expression of let-7a; and the levels of caspase-3 and p53 proteins in HeLa cells were determined by Western blot analysis. Differences between the mean values of multiple groups were analyzed by one-way analysis of variance or Student's t test. P < 0.05 was considered statistically significant. Results: Compared to control group, obvious morphological changes were observed in the SeO2 group. SeO2 significantly (P < 0.05) inhibited cell proliferation and viability in a dose-dependent manner (7.5 μmol/L, 15 μmol/L, and 30 μmol/L of SeO2), as demonstrated by MTT assay. FCM analysis revealed that SeO2, dose dependently, increased the apoptotic rate of the treated cells. SeO2 up-regulated the caspase-3 and p53 levels which peaked at the concentration of 7.5 μmol/L in HeLa cells. It also significantly (P < 0.05) induced the expression of apoptosis-related microRNA let-7a in both cell lines, which reached its peak at the concentration of 7.5 μmol/L. Conclusion: SeO2 showed anti-tumor properties via apoptotic pathway by up-regulating the expressions of let-7a, as well as caspase-3 and p53 in cervical cancer cells

Small Extracellular Vesicles Secreted by iPSC-Derived MSCs Ameliorate Pulmonary Inflammation and Lung Injury Induced by Sepsis through Delivery of miR-125b-5p

Background. Sepsis-induced acute lung injury is a common critical illness in intensive care units with no effective treatment is currently available. Small extracellular vesicles, secreted by mesenchymal stem cells (MSCs), derived from human-induced pluripotent stem cells (iMSC-sEV), possess striking advantages when incorporated MSCs and iPSCs, which are considered extremely promising cell-free therapeutic agents. However, no studies have yet been conducted to systemically examine the effects and underlying mechanisms of iMSC-sEV application on attenuated lung injury under sepsis conditions. Method. iMSC-sEV were intraperitoneally administered in a rat septic lung injury model induced by cecal ligation and puncture (CLP). The efficacy of iMSC-sEV was assessed by histology, immunohistochemistry, and pro-inflammatory cytokines of bronchoalveolar lavage fluid. We also evaluated the in vitro effects of iMSC-sEV on the activation of the inflammatory response in alveolar macrophages (AMs). Small RNA sequencing was utilized to detect changes in the miRNA expression profile in lipopolysaccharide (LPS)-treated AMs after iMSC-sEV administration. The effects of miR-125b-5p on the function of AMs were studied. Results. iMSC-sEV were able to attenuate pulmonary inflammation and lung injury following CLP-induced lung injury. iMSC-sEV were internalized by AMs and alleviated the release of inflammatory factors by inactivating the NF-κB signaling pathway. Moreover, miR-125b-5p showed a fold-change in LPS-treated AMs after iMSC-sEV administration and was enriched in iMSC-sEV. Mechanistically, iMSC-sEV transmitted miR-125b-5p into LPS-treated AMs to target TRAF6. Conclusion. Our findings demonstrated that iMSC-sEV treatment protects against septic lung injury and exerts anti-inflammatory effects on AMs at least partially through miR-125b-5p, suggesting that iMSC-sEV may provide a novel cell-free strategy for the treatment of septic lung injury

Generation of special autosomal dominant polycystic kidney disease iPSCs with the capability of functional kidney-like cell differentiation

Abstract Background Human induced pluripotent stem cells (iPSCs) have been verified as a powerful cell model for the study of pathogenesis in hereditary disease. Autosomal dominant polycystic kidney disease (ADPKD) is caused by mutations of PKD or non-PKD genes. The pathogenesis of ADPKD remains unexplored because of the lack of a true human cell model. Methods Six ADPKD patients and four healthy individuals were recruited as donors of somatic cells from a Chinese ADPKD family without mutations of the PKD genes but carrying SAMSN1 gene deletion. The ADPKD-iPSCs were generated from somatic cells and were induced into kidney-like cells (KLCs) by a novel three-step method involving cytokines and renal epithelium growth medium. Furthermore, we analyzed functional properties of these KLCs by water transportation and albumin absorption assays. Results We successfully generated iPSCs from ADPKD patients and differentiated them into KLCs that showed morphological and functional characteristics of human kidney cells. Further, we also found that ADPKD-iPSC-KLCs had a significantly higher rate of apoptosis and a significantly lower capacity for water transportation and albumin absorption compared to healthy sibling-derived differentiated KLCs. Furthermore, knockdown of SAMSN1 in control iPSCs may attenuate differentiation and/or function of KLCs. Conclusions These data show that we have created the first iPSCs established from ADPKD patients without mutations in the PKD genes, and suggest that the deletion mutation of SAMSN1 might be involved in the differentiation and/or function of KLCs. ADPKD-iPSC-KLCs can be used as a versatile model system for the study of kidney disease

Additional file 2: Figure S2. of Generation of special autosomal dominant polycystic kidney disease iPSCs with the capability of functional kidney-like cell differentiation

The Sanger sequencing analysis for PKD in a Chinese ADPKD family. (a): The novel missense mutation c.17 G > A, p.Arg6His in PKD2 was predicted by three program. (b): The list of all ten persons analyzed for the mutations. (c): The real sequencing pictures of all ten individuals in this family. (JPG 4280 kb

Additional file 1: Figure S1. of Generation of special autosomal dominant polycystic kidney disease iPSCs with the capability of functional kidney-like cell differentiation

The additional characterization analysis for ADPKD-iPSC and KLCs. (a): The timeline and culture conditions of induction of fibroblasts to iPSCs. Lower panel; phase contrast microscopy showing each of the three major steps. Bar = 100um. (b): AP staining for stemness of stem cells in iPSC lines. Bar = 100um. (c): Immunofluorescence photomicrographs showing primary cilia (arrow head) in KCLs were generated from iPSCs. Bar = 5um. (d): Water transportation assays were carried out between HK2 positive cells and KCLs. Data are represented as mean ± standard deviation from three independent sets of experiments. (e): The podocyte was used as a positive control and absorbed rhodamine-albumin. Bar = 25um. (JPG 1230 kb