Exosome Marker Proteins of Tumor-Associated Fibroblasts and Exosome-Derived miR-92a-3p Act as Potential Biomarkers for Liver Cancer.

Many recent studies have shown that microRNAs (miRNAs) in exosomes can be absorbed by nearby or distant cells, and the abnormal expression of these exosomal miRNAs is associated with most pathological progresses. In this study, we explored the diagnostic value of exosome marker proteins and exosome-derived miR-92a-3p in liver cancer. The clinicopathological data of 60 patients with liver cancer admitted to Tanghan Gongren Hospital from October 2017 to October 2019 were collected. Tumor tissue and adjacent tissue were collected during surgery. Quantitative reverse transcription polymerase chain reaction and Western blot were used to detect the expression levels of miR-92a-3p in exosomes of fibroblasts and tumor tissue, and exosome marker proteins. In liver cancer tissue and fibroblast exosomes, the expression of miR-92a-3p was significantly increased. The receiver operator characteristic curve of the expression level of miR-92a-3p in exosomes and tissue showed that the area under the curve was 0.906 and 0.911, respectively. HSP70 and CD63 were highly expressed in the tissue of liver cancer and fibroblast exosomes. miR-92a-3p was positively correlated with HSP70 and CD63 in the exosomes of liver cancer fibroblasts. In addition, miR-92a-3p and exosome marker proteins (HSP70 and CD63) were highly expressed in tumors with a diameter greater than 5 cm, and were higher in liver cancer patients with BCLC stage B/C. Tumor fibroblast-derived exosome marker proteins and miR-92a-3p have good diagnostic value in liver cancer, indicating that they may be new diagnostic markers for liver cancer.

Identification and construction of a 13-gene risk model for prognosis prediction in hepatocellular carcinoma patients.

We attempted to screen out the feature genes associated with the prognosis of hepatocellular carcinoma (HCC) patients through bioinformatics methods, to generate a risk model to predict the survival rate of patients. Gene expression information of HCC was accessed from GEO database, and differentially expressed genes (DEGs) were obtained through the joint analysis of multi-chip. Functional and pathway enrichment analyses of DEGs indicated that the enrichment was mainly displayed in biological processes such as nuclear division. Based on TCGA-LIHC data set, univariate, LASSO, and multivariate Cox regression analyses were conducted on the DEGs. Then, 13 feature genes were screened for the risk model. Also, the hub genes were examined in our collected clinical samples and GEPIA database. The performance of the risk model was validated by Kaplan-Meier survival analysis and receiver operation characteristic (ROC) curves. While its universality was verified in GSE76427 and ICGC (LIRI-JP) validation cohorts. Besides, through combining patients' clinical features (age, gender, T staging, and stage) and risk scores, univariate and multivariate Cox regression analyses revealed that the risk score was an effective independent prognostic factor. Finally, a nomogram was implemented for 3-year and 5-year overall survival prediction of patients. Our findings aid precision prediction for prognosis of HCC patients.

miR-9-5p facilitates hepatocellular carcinoma cell proliferation, migration and invasion by targeting ESR1.

The study aimed to explore the relationship between miR-9-5p and ESR1, and clarify the underlying functional mechanism in the occurrence and development of hepatocellular carcinoma (HCC). Expression data including miRNAs and mRNAs of HCC downloaded from TCGA database were processed for differential analysis, and corresponding clinical data were collected for survival analysis to identify the target miRNA miR-9-5p. Bioinformatics databases were applied for predicting downstream target mRNAs of miR-9-5p. qRT-PCR was used to evaluate expression of miR-9-5p. Western blot was used to detect protein expression of ESR1. MTT, wound healing assay and Transwell assay were used to detect HCC cell proliferation, migration and invasion, respectively. Dual-luciferase reporter gene assay was used to identify the targeting relationship between miR-9-5p and ESR1. Research suggested that miR-9-5p was highly expressed in HCC cells but ESR1 was poorly expressed. Overexpression of miR-9-5p could improve the proliferation, invasion and migration of cells. Dual-luciferase reporter assay showed that ESR1 was the downstream target of miR-9-5p in HCC. Overexpression of miR-9-5p markedly reduced ESR1 mRNA and protein levels in HCC cells, whereas inhibition of miR-9-5p expression produced the contrary results. Silencing ESR1 could noticeably reverse the effect of miR-9-5p knockdown on the proliferation, migration and invasion of HCC cells. As an oncogene, miR-9-5p fostered the proliferation, migration and invasion of HCC cells by targeting and inhibiting ESR1 expression.

m5C RNA methyltransferase-related gene NSUN4 stimulates malignant progression of hepatocellular carcinoma and can be a prognostic marker.

Hepatocellular carcinoma (HCC) is a cancer with relatively high mortality, yet little attention has been devoted for related prognostic biomarkers. This study analyzed differential expression of m5C RNA methyltransferase-related genes in normal samples and tumors samples in TCGA-LIHC using Wilcoxon test. K-means consensus clustering analysis was implemented to subdivide samples. Independent prognostic factors were screened by univariate and multivariate Cox regression analyses. KEGG pathway enrichment analysis was performed on the screened independent prognostic factor using GSEA tools. qPCR was conducted to test mRNA expression of key m5C RNA methyltransferase-related genes in tissues and cells. There were 7 m5C RNA methyltransferase-related genes (NOP2, NSUN4, etc.) differentially expressed in HCC tumor tissues. HCC samples were classified into 3 subgroups through clustering analysis according to the expression mode of m5C RNA methyltransferase-related genes. It was also discovered that patients in different subgroups presented significant differences in survival rate and distribution of grade. Additionally, NOP2, NSUN4 and NSUN5 expression notable varied in different grades. Through regression analyses combined with various clinical pathological factors, it was displayed that NSUN4 could work as an independent prognostic factor. KEGG analysis showed that NSUN4 mainly enriched in signaling pathways involved in ADHERENS JUNCTION, RNA DEGRADATION, MTOR SIGNALING PATHWAY, COMPLEMENT and COAGULATION CASCADES. As examined by qPCR, NSUN4 was conspicuously upregulated in HCC patient's tissues and cells. Altogether, our study preliminarily developed a novel biomarker that could be independently used in prognosis of HCC, which may provide a new direction for the study of related molecular mechanism or treatment regimen.

MiR-18a-5p Facilitates Progression of Hepatocellular Carcinoma by Targeting CPEB3.

Objective: To explore the function of the miR-18a-5p/CPEB3 axis in regulating the occurrence of hepatocellular carcinoma (HCC). Methods: Differentially expressed miRNAs and mRNAs were acquired by bioinformatics analysis. qRT-PCR was used for miR-18a-5p and CPEB3 mRNA expression detection. Cell functional assays were implemented to examine the biological functions of HCC cells. The binding relationship between miR-18a-5p and CPEB3 was verified by a dual luciferase assay. Results: In HCC, miR-18a-5p was remarkably highly expressed, while CPEB3 was markedly lowly expressed. HCC cell progression was facilitated after cells transfecting miR-18a-5p mimic, whereas silencing miR-18a-5p caused the opposite result. Overexpressing CPEB3 could restore promoting effect of miR-18a-5p on the growth of HCC cells. Conclusion: Oncogene miR-18a-5p accelerates malignant phenotype by suppressing CPEB3. MiR-18a-5p/CPEB3 axis in HCC identified in this study provides a new target for HCC treatment.

mir-182-5p Regulates Cell Growth of Liver Cancer via Targeting RCAN1.

Regulator of calcineurin 1 (RCAN1) is an endogenous protein that is involved in the regulation of the occurrence and progression of a variety of cancers, but currently, people know little about its potential mechanism. This study investigated the function and mechanism of RCAN1 and miR-182-5p in liver cancer cells. In this study, reliable data demonstrated that RCAN1 suppressed cell proliferation, migration, invasion, and cell cycle progression of liver cancer. Additionally, the expression of RCAN1 was noted to be regulated by its upstream regulator miR-182-5p, and miR-182-5p was prominently highly expressed in liver cancer cells. Based on this, it was further proved through cell experiments that miR-182-5p facilitated cell growth of liver cancer through RCAN1 downregulation, showing that RCAN1 may be a fresh biomarker and target for diagnosis and treatment of liver cancer.

MiR-92a-3p Promotes the Malignant Progression of Hepatocellular Carcinoma by Mediating the PI3K/AKT/mTOR Signaling Pathway.

BACKGROUND: As one of the most common cancers globally, hepatocellular carcinoma (HCC) usually has a poor prognosis. Many HCC patients are usually diagnosed at advanced stages. Therefore, new potential biomarkers for the diagnosis and prognosis of HCC are urgently needed. More and more studies have shown that miR-92a-3p can regulate the occurrence and development of various cancers, but its clinical significance and molecular mechanism in HCC are still elusive. Here, we tried to clarify the regulatory mechanism of miR-92a-3p in HCC. METHODS: In this study, we conducted qRT-PCR and revealed that miR-92a-3p was notably upregulated in HCC cells. MTT, flow cytometry, wound healing, Transwell invasion assays and western blot were conducted to uncover that overexpressed miR-92a-3p could boost the proliferation, migration, invasion and epithelial-mesenchymal transition (EMT) of HCC cells while inhibiting cell apoptosis. In addition, the proteins associated with the PI3K/AKT/mTOR pathway were also detected by western blot. RESULTS: It was suggested that miR-92a-3p could activate the PI3K/AKT/mTOR signaling pathway. CONCLUSION: These results suggest that miR-92a-3p plays a tumor-promoting role in HCC and may be a potential biomarker for the diagnosis and prognosis of HCC.

MiR-452-5p mediates the proliferation, migration and invasion of hepatocellular carcinoma cells via targeting COLEC10.

Objective: This study explored the potential function of miR-452-5p in hepatocellular carcinoma (HCC) and clarified the mechanism underlying HCC progression. Materials & methods: Real-time quantitative PCR was used to detect miR-452-5p and COLEC10 mRNA expression in HCC, western blot was performed to test COLEC10 protein expression. The regulatory mechanism of miR-452-5p/COLEC10 in HCC cells was explored using CCK-8, wound healing assay, Transwell and dual-luciferase reporter assay. Results: MiR-452-5p was greatly upregulated in HCC cells, and it served as an oncogene playing an active role in HCC cell proliferation, migration and invasion. COLEC10 was identified as the target of miR-452-5p in HCC attenuating the promoting effect of miR-452-5p on HCC cells upon overexpression. Conclusion: MiR-452-5p can promote the progression of HCC via targeting COLEC10.

Separation distance dependent fluorescence enhancement of fluorescein isothiocyanate by silver nanoparticles.

Nanocomposites consisting of a metal core, a silica-spacer shell with controlled thickness, and a dye-labelled shell were synthesized and separation distance dependent fluorescence enhancement of fluorescein isothiocyanate by silver nanoparticles was studied; the results indicated an optimum enhancement of 4.8 times with a spacer shell thickness of 21 nm.

Controlled synthesis of Y-junction polyaniline nanorods and nanotubes using in situ self-assembly of magnetic nanoparticles.

Novel Y-junction polyaniline (PANI) nanorods and nanotubes (40-100 nm in diameter and a few micrometers in length) were selectively prepared, for the first time, using in situ self-assembly of water-soluble Fe3O4 nanoparticles coated with polyethylene glycol(5) nonylphenyl ether (NP5) and cyclodextrin (CD) as templates and pH control in an aqueous medium. The morphology of Fe3O4 nanoparticles and the PANI nanostructures (rods and tubes) was determined by TEM and the effects of reaction conditions on the morphology of PANI nanostructures were also studied. Characterization experiments including FTIR and X-ray diffraction were carried out to study the chemical and electronic structures of the PANI nanostructures.

Formation of ordered arrays of oriented polyaniline nanoparticle nanorods.

We report the preparation of ordered polyaniline (PANI) nanorod arrays in an aqueous medium. The oriented PANI nanorods (80-400 nm in diameter and 8-15 mum in length) were synthesized in the presence of hydrophilic Allura Red AC (ARAC) as the structure-directing agent and ammonium persulfate as an oxidant in HCl solution. The morphologies of the oriented PANI nanoparticle nanorods were confirmed by scanning electron microscopy (SEM) and transmission electron microscopy images, and the effect of reaction conditions on the morphology of PANI nanostructures was also studied. On the basis of the result obtained from small-angle X-ray scattering, we propose that rodlike micelle arrays of ARAC-aniline are responsible for directing the formation of oriented PANI nanoparticle nanorods. SEM images and the data analysis of static and dynamic light scattering give supportive evidence to the formation of the PANI nanoparticle nanorods by an elongation process. The chemical and electronic structures of the PANI nanorods were also studied by Fourier transform IR and UV-vis spectrometries, respectively.

Preparation, characterization, and drug release behaviors of drug nimodipine-loaded poly(epsilon-caprolactone)-poly(ethylene oxide)-poly(epsilon-caprolactone) amphiphilic triblock copolymer micelles.

Amphiphilic triblock copolymers, poly(epsilon-caprolactone)-poly(ethylene oxide)-poly(epsilon-caprolactone) (PCL-PEO-PCL), were synthesized by ring opening polymerization of epsilon-caprolactone initiated with the hydroxyl functional groups of poly(ethylene glycol) at both ends of the chain. The micelles composed of this type of copolymer had such a structure that both ends of the PEO chain were anchored to the micelle. The critical micelle concentration of the block copolymer in distilled water was determined by a fluorescence probe technique using pyrene. As the hydrophobic components of the block copolymer increased, the critical micelle concentration value decreased. To estimate the feasibility as novel drug carriers, the block copolymer micelles were prepared by precipitation of polymer from acetone solution into water. From the observation of transmission electron microscopy, the micelles exhibited a spherical shape. Nimodipine was incorporated into the hydrophobic inner core of micelles as a lipophilic model drug to investigate the drug release behavior. The PEO/PCL ratio of copolymer was a main factor in controlling micelle size, drug-loading content, and drug release behavior. As PCL weight ratio increased, the micelle size and drug-loading content increased, and the drug release rate decreased.

Microfluidic sensing devices employing in situ-formed liquid crystal thin film for detection of biochemical interactions.

Although biochemical sensing using liquid crystals (LC) has been demonstrated, relatively little attention has been paid towards the fabrication of in situ-formed LC sensing devices. Herein, we demonstrate a highly reproducible method to create uniform LC thin film on treated substrates, as needed, for LC sensing. We use shear forces generated by the laminar flow of aqueous liquid within a microfluidic channel to create LC thin films stabilized within microfabricated structures. The orientational response of the LC thin films to targeted analytes in aqueous phases was transduced and amplified by the optical birefringence of the LC thin films. The biochemical sensing capability of our sensing devices was demonstrated through experiments employing two chemical systems: dodecyl trimethylammonium bromide (DTAB) dissolved in an aqueous solution, and the hydrolysis of phospholipids by the enzyme phospholipase A(2) (PLA(2)).

Fabrication of polymeric hollow nanospheres, hollow nanocubes and hollow plates.

A facile strategy for fabricating polypyrrole-chitosan (PPy-CS) hollow nanostructures with different shapes (sphere, cube and plate) and a wide range of sizes (from 35 to 600 nm) is described. These hollow structures have been fabricated using silver bromide as a single template material for polymer nucleation and growth. PPy-CS hollow nanostructures are formed by reaction with an etching agent to remove the core. These hollow nanostructures have been extensively characterized using various techniques such as TEM, FT-IR, UV-vis, and XRD.

Facile fabrication of AgCl@polypyrrole-chitosan core-shell nanoparticles and polymeric hollow nanospheres.

A one-step sequential method for preparing AgCl@polypyrrole-chitosan core-shell nanoparticles and subsequently the formation of polypyrrole-chitosan hollow nanospheres is reported. The formation of the core and the shell is performed in one reaction medium almost simultaneously. Transmission electron microscopy (TEM) images show the presence of core-shell nanoparticles and hollow nanospheres. Ultraviolet-visible (UV-vis) studies reveal that AgCl was formed first followed by polypyrrole. X-ray diffration (XRD) and UV-vis studies show that AgCl was present in the core-shell nanoparticles and could be removed completely from the core.

[Attachment and growth of cultured fibroblast cells on chitosan/PHEA-blended hydrogels].

The chitosan/PHEA-blended hydrogels were prepared from PHEA and chitosan in various blend ratios. The water contents of the hydrogels were in the range of 50%-80% (wt). The attachment and growth of fibroblast cells(L929) on the hydrogels were studied. The results indicated the PHEA content in hydrogels has great effect on cell attachment but has little effect on the growth of L929 cells.