Proteomic analysis of human umbilical cord serum exosomes using mass spectrometry and preliminary study of their biological activities in liver cancer cell lines.

Exosomes are membranous extracellular vesicles 50-100 nm in size, which are involved in cellular communication via the delivery of proteins, lipids and RNA. Emerging evidence shows that exosomes play a critical role in cancer. It has recently been revealed that maternal and umbilical cord serum (UCS)-derived exosomes may enhance endothelial cell proliferation and migration. However, the role of exosomes isolated from the human umbilical cord in cancer development has not been investigated. To explore the potential differences in the composition and function of proteins from umbilical serum exosomes (UEs) and maternal serum exosomes, a proteomic analysis of exosomes was conducted using mass spectrometry and bioinformatics. Moreover, Cell Counting Kit-8 assays and flow cytometry were used to study the biological effects of UEs on liver cancer cell lines. The present study demonstrated that UCS was enriched with proteins involved in extracellular matrix-receptor interactions, which may be closely related to cell metastasis and proliferation. The findings further indicated that exosomes derived from human umbilical serum could inhibit the viability and induce apoptosis of liver cancer cells. This suggests that UCS-derived exosomes may represent potential leads for the development of biotherapy for liver cancer.

Tubulin-binding peptide RR-171 derived from human umbilical cord serum displays antitumor activity against hepatocellular carcinoma via inducing apoptosis and activating the NF-kappa B pathway.

OBJECTIVES: Hepatocellular carcinoma (HCC) still presents a high incidence of malignant tumours with poor prognosis. There is an urgent need for new therapeutic agents with high specificity, low toxicity and favourable solubility for the clinical treatment of HCC. MATERIALS AND METHODS: The bioactivity of human umbilical cord serum was investigated by proteomics biotechnology and a primitive peptide with certain biological activity was identified. The antitumour effect of RR-171 was detected by cell viability assay in vitro, and determined by subcutaneous xenograft models assay and miniPDX assay in vivo. Pull-down experiments were conducted to identify the potential targeting proteins of RR-171. Immunofluorescence assay and tubulin polymerization assay were conducted to explore the relationship between RR-171 and α-tubulin. Fluorescence imaging in xenograft models was used to explore the biodistribution of RR-171 in vivo. A phosphospecific protein microarray was performed to uncover the underlying signalling pathway by which RR-171 induces tumour cell death. RESULTS: The results indicated that RR-171 could be effective in the treatment of HCC in vivo and in vitro. RR-171 could aggregate significantly in solid tumours and had no obvious systemic toxicity in vivo. RR-171 could interact with α-tubulin and activate the NF-Kappa B pathway in HCC cells. CONCLUSIONS: Taken together, RR-171 exhibited significant antitumour activity against HCC in vivo and in vitro and could potentially be used in the clinical application of HCC.

Potent antitumor activity of a glutamyltransferase-derived peptide via an activation of oncosis pathway.

Hepatocellular carcinoma (HCC) still presents poor prognosis with high mortality rate, despite of the improvement in the management. The challenge for precision treatment was due to the fact that little targeted therapeutics are available for HCC. Recent studies show that metabolic and circulating peptides serve as endogenous switches for correcting aberrant cellular plasticity. Here we explored the antitumor activity of low molecular components in human umbilical serum and identified a high abundance peptide VI-13 by peptidome analysis, which was recognized as the part of glutamyltransferase signal peptide. We modified VI-13 by inserting four arginines and obtained an analog peptide VI-17 to improve its solubility. Our analyses showed that the peptide VI-17 induced rapid context-dependent cell death, and exhibited a higher sensitivity on hepatoma cells, which is attenuated by polyethylene glycol but not necrotic inhibitors such as z-VAD-fmk or necrostatin-1. Morphologically, VI-17 induced cell swelling, blebbing and membrane rupture with release of cellular ATP and LDH into extracellular media, which is hallmark of oncotic process. Mechanistically, VI-17 induced cell membrane pore formation, degradation of α-tubulin via influx of calcium ion. These results indicated that the novel peptide VI-17 induced oncosis in HCC cells, which could serve as a promising lead for development of therapeutic intervention of HCC.

Dysbindin promotes pancreatic ductal adenocarcinoma metastasis by activating NF-κB/MDM2 via miR-342-3p.

Pancreatic ductal adenocarcinoma (PDAC) is one of the most invasive solid tumours and has the highest cancer-related mortality rate. Despite intense investigation, the molecular mechanisms underlying the invasiveness and aetiology of PDAC remain elusive. MicroRNAs (miRNAs) are key regulators of tumour cell plasticity, but their roles in PDAC metastasis have not been characterized. Our early studies showed that dysbindin protein levels are elevated in PDAC patients compared with control individuals and that dysbindin upregulation elicits PDAC cell proliferation via the PI3K pathway. Here, we show that dysbindin promoted PDAC metastasis via the NF-κB/MDM2 signalling axis. Increased dysbindin levels correlated with aggressive features in PDAC, and the overexpression of dysbindin significantly promoted PDAC metastasis and invasion in vitro and in vivo. Surprisingly, dysbindin was identified as a direct target of miR-342-3p, which promotes NF-κB activation and PDAC metastasis. Thus, dysbindin-mediated NF-κB activation via miR-342-3p represents a context-dependent switch that enables PDAC cell proliferation and metastasis. Our data suggest that dysbindin and miR-342-3p are potential leads for the development of targeted therapy for PDAC.

Insulin-like growth factor-1 and non-alcoholic fatty liver disease: a systemic review and meta-analysis.

AIM: The prevalence of non-alcoholic fatty liver disease (NAFLD) is rapidly increasing worldwide. A number of researchers have studied the relationship between Insulin-like growth factor-1(IGF-1) and NAFLD. However, the results are controversial. This meta-analysis, aimed to systemically evaluate the correlation between IGF-1 and NAFLD. METHODS: We searched for four online databases: PubMed, Web of Science, Embase and CNKI up to Feb 2018. We then applied a random-effects model to evaluate the overall effect sizes by calculating Standard mean difference (SMD) and its 95% confidence intervals (CIs). RESULTS: Twelve articles were included in this meta-analysis. The pooled analysis showed that the level of IGF-1 in the control group was significantly higher than that in the NAFLD group. (SMD: 1.00, 95% CI: 0.54-1.46, P < 0.00001). However, significant heterogeneity was discovered among the included studies (P < 0.00001, I2 = 96%). Then a series of subgroup analyses were performed. Compared to the nonalcoholic steatohepatitis (NASH) group, the level of IGF-1 was significantly higher in the Non- or probable-NASH group (SMD: 1.42, 95% CI: 0.25-2.58, P = 0.02). The level of IGF-1 in patients with increased insulin resistance (SMD: 0.49; 95% CI: 0.36-0.63; P < 0.00001) and high Body Mass Index (SMD: 0.50; 95% CI: 0.22-0.79; P < 0.05) were significantly lower than healthy control. In addition, the same conclusion were found in studies carried out in Asia and Europe (Asia: SMD: 0.69, 95% CI: -0.29-1.66, P = 0.17; Europe: SMD: 0.89, 95% CI: 0.41-1.38, P < 0.05). CONCLUSION: The level of IGF-1 is down-regulated in NAFLD patients compared to healthy controls, suggesting that IGF-1 might be used as a potential biomarker and therapeutic target for NAFLD.