Adipose-derived stem cell-derived microvesicle-released miR-210 promoted proliferation, migration and invasion of endothelial cells by regulating RUNX3

<p>The potential mechanism of miRNA released from adipose-derived stem cell (ADSC)-derived micro vesicle (MV) onthe modulation of proliferation, migration and invasion of endothelial cells were explored. In this study, miR-210 level was detected by qT-PCR. Alix, VEGF and RUNX3 expressions were detected by Western blot. The proliferation, migration and invasion of human umbilical vein endothelial cells (HUVECs) were observed by MTT assay and Transwell assay. Luciferase reporter gene assay was conducted to validate the targeting activity of MVs-released miR-210 on RUNX3. We found hypoxia significantly increased the expression of MVs-released miR-210. MVs released from ADSCsin hypoxic group significantly promoted the proliferation, migration and invasion of HUVECs. Overexpression of miR-210 significantly upregulated VEGF expression, and promoted the proliferation, migration and invasion of HUVECs. Besides, RUNX3 was identified as the direct of miR-210 in HUVECs. Overexpression of miR-210 decreased RUNX3 expression and promoted the proliferation, migration and invasion of HUVECs, while overexpression of RUNX3 inhibited these promotion effects. In vivo experiment showed that MVs derived from ADSCs under hypoxia increased miR-210 level and capillary density, and inhibition of miR-210 decreased capillary density. We also found MVs downregulated RUNX3 expression, and inhibition of miR-210 upregulated RUNX3 expression. Therefore, miR-210 released from ADSCs-derived MVs promoted proliferation, migration and invasion of HUVECs by targeting RUNX3, which revealed one of the mechanisms of ADSCs-derived MVs on the promotion of proliferation, migration and invasion of HUVECs.</p> <p><b>Abbreviations</b>: ADSC, adipose-derived stem cell; MV, micro vesicle; HUVECs, human umbilical vein endothelial cells; RUNX3, Runtrelatedtranscription factor-3</p

Enhancing Glioblastoma-Specific Penetration by Functionalization of Nanoparticles with an Iron-Mimic Peptide Targeting Transferrin/Transferrin Receptor Complex

Treatment of glioblastoma (GBM) remains to be the most formidable challenge because of the hindrance of the blood–brain barrier (BBB) along with the poor drug penetration into the glioma parenchyma. Nanoparticulate drug delivery systems (DDS) utilizing transferrin (Tf) as the targeting ligand to target the glioma-associated transferrin receptor (TfR) had met the problem of loss of specificity in biological environment due to the high level of endogenous Tf. Here we conjugated CRT peptide, an iron-mimicry moiety targeting the whole complex of Tf/TfR, to poly­(ethylene glycol)-poly­(l-lactic-<i>co</i>-glycolic acid) nanoparticles (CRT-NP), to open a new route to overcome such obstacle. High cellular associations, advanced transport ability through the BBB model, and penetration in 3-dimensional C6 glioma spheroids <i>in vitro</i> had preliminarily proved the advantages of CRT-NP over Tf-nanoparticle conjugates (Tf-NP). Compared with Tf-NP, NP, and Taxol, paclitaxel-loaded CRT-NP (CRT-NP-PTX) displayed a superior antiproliferation effect on C6 glioma cells and stronger inhibitory effect on glioma spheroids. Favored pharmacokinetics behavior and enhanced accumulation in glioma foci was observed, together with a much deeper distribution pattern in glioma parenchyma compared with unmodified nanoparticles and Tf-NP. Eventually, mice treated with CRT-NP-PTX showed a remarkably prolonged median survival compared to those treated with Taxol, NP, or Tf-NP. In conclusion, the modification of CRT to nanoparticles holds great promise for enhancement of antiglioma therapy

Relationship between the TT and the clinical characteristics in 204 patients with ESCC.

<p>Bold italics indicate significant differences (<i>p</i> < 0.05). Mean ± SD, Mean ± standard deviation; TT, Thrombin Time.</p><p>*P values were calculated using unpaired Student’s t-tests or one-way ANOVA, <i>p</i> <0.05 indicated significant differences.</p><p>**P values were calculated using the chi-squared test (χ² test), <i>p</i> <0.05 indicated significant differences.</p><p>^♯TNM denotes tumor-node-metastasis.</p><p>Relationship between the TT and the clinical characteristics in 204 patients with ESCC.</p

Main Clinical Characteristics of Patients Group According to Coagulation Parameter Levels.

<p>Bold italics indicate significant differences (<i>p</i> < 0.05).</p><p>*Moderately and well differentiated histologic types were classified as differentiated carcinoma; Low levels of differentiation were defined as undifferentiated carcinoma.</p><p>Main Clinical Characteristics of Patients Group According to Coagulation Parameter Levels.</p

Prognostic significance of serum TT in ESCC.

<p>The patients were categorized into a low TT group and a normal TT group according to the media value (17.2 sec). The five-year overall survival rate was calculated using the Kaplan-Meier method and analyzed with the log-rank test. A high TT level was a favorable prognostic factor in the entire ESCC cohort (a), T3–T4 subgroup (c), N1–N2 subgroup (e), and M0 subgroup (g).</p

The serum coagulation tests results in patients with ESCC and healthy controls.

<p>The serum coagulation tests results in patients with ESCC and healthy controls.</p

Univariate and multivariate cox hazards analysis for overall survival in 204 patients with ESCC.

<p>HR, Hazard ratio; 95% CI, 95% confidence interval; PT, Prothrombin Time; PTA%, Prothrombin Time Activity; INR, International Normalized Ratio; APTT, Activated Partial Thromboplastin Time; TT, Thrombin Time.</p><p>*Moderately and well differentiated histologic types were classified as differentiated carcinoma; Low levels of differentiation were defined as undifferentiated carcinoma.</p><p>**Cox hazard regression model.</p><p>^♯TNM denotes tumor-node-metastasis.</p><p>Univariate and multivariate cox hazards analysis for overall survival in 204 patients with ESCC.</p

Effect of PHB knockdown on the morphology of mitochondria in 3T3-L1 cells.

<p>3T3-L1 cells were transfected with siRNA targeting PHB1 or PHB2, and cultured for three days. siControl was used as the control. <b>A.</b> The cristae morphology of mitochondria (arrows) in 3T3-L1 cells, transfected with the indicated siRNAs, was assessed by transmission electron microscopy (TEM) at 80 kV acceleration voltage and 50 k magnification. Scale bar, 100 nm. <b>B.</b> The siRNA-transfected 3T3-L1 cells were subject to adipogenic induction for 7 days. The cells at day 0 and day 7 were stained with MitoTracker Red. The mitochondrial morphology was analyzed with a Confocal Microscopy System. Scale bar, 10 µm. High-power magnification insets are included to emphasize the comparison of tubular mitochondria to fragmented ones. The bar graph represents the percentage of cell populations with fragmented mitochondria. **<i>p</i><0.01 compared to siControl at day 0; ^##<i>p</i><0.01 compared to siControl at day 7.</p

Content of mitochondrial PHBs and mtDNA in 3T3-L1 cells pre- and post-adipogenesis.

<p><b>A, B and C.</b> Over-confluent 3T3-L1 cells (Day 0) were induced for adipocyte differentiation for 7 days. <b>A,</b> PHB1 (green) or <b>B,</b> PHB2 (green) was detected by using immunocytochemistry. CytC (red) was used as the mitochondrial marker. Bar = 10 µm. <b>C.</b> The levels of PHB1 and PHB2 in isolated mitochondria were detected using the immunoblotting analysis. The mitochondrial marker porin, also called the voltage dependent anion channel (VDAC), was used as a loading control. <b>D.</b> 3T3-L1 cells were transfected with indicated siRNAs and cultured for three days. The relative mtDNA content was evaluated by a ratio of the DNA level of mitochondrial Complex I to the DNA level of nuclear 18SrRNA. The relative mtDNA content in 3T3-L1 cells transfected with siControl was set to 1. *p<0.05, **p<0.01 compared to siControl (Day 0). ^#p<0.05, ^##p<0.01 compared to siControl (Day 7).</p

Nanoparticles Coated with Neutrophil Membranes Can Effectively Treat Cancer Metastasis

The dissemination, seeding, and colonization of circulating tumor cells (CTCs) serve as the root of distant metastasis. As a key step in the early stage of metastasis formation, colonization of CTCs in the (pre-)­metastatic niche appears to be a valuable target. Evidence showed that inflammatory neutrophils possess both a CTC- and niche-targeting property by the intrinsic cell adhesion molecules on neutrophils. Inspired by this mechanism, we developed a nanosize neutrophil-mimicking drug delivery system (NM-NP) by coating neutrophils membranes on the surface of poly­(latic-<i>co</i>-glycolic acid) nanoparticles (NPs). The membrane-associated protein cocktails on neutrophils membrane were mostly translocated to the surface of NM-NP <i>via</i> a nondisruptive approach, and the biobinding activity of neutrophils was highly preserved. Compared with uncoated NP, NM-NP exhibited enhanced cellular association in 4T1 cell models under shear flow <i>in vitro</i>, much higher CTC-capture efficiency <i>in vivo</i>, and improved homing to the premetastatic niche. Following loading with carfilzomib, a second generation of proteasome inhibitor, the NM-NP-based nanoformulation (NM-NP-CFZ) selectively depleted CTCs in the blood, prevented early metastasis and potentially inhibited the progress of already-formed metastasis. Our NP design can neutralize CTCs in the circulation and inhibit the formation of a metastatic niche