The oncogenetic role of microRNA-31 as a potential biomarker in oesophageal squamous cell carcinoma,”

A B S T R A C T miR-31 (microRNA-31) is frequently altered in numerous cancers. The aim of the present study was to investigate the role of miR-31 in ESCC (oesophageal squamous cell carcinoma). We measured miR-31 in 45 paired ESCC tissues and 523 serum samples using real-time RT (reverse transcription)-PCR. The serum samples were divided into a discovery group (120 ESCCs and 121 normal controls), a validation group (81 ESCCs and 81 controls), and a final group comprising six other common tumours (colorectal, liver, cervical, breast, gastric and lung cancers; total n = 120). A Mann-Whitney U test and Wilcoxon matched-pairs test were used for the statistics. miR-31 was up-regulated in 77.8 % of the ESCC tissues. Serum miR-31 levels in ESCC patients were significantly higher than in normal controls (P < 0.001). Patients with high-levels of serum miR-31 also had a poorer prognosis in relapse-free survival (P = 0.001) and tumour-specific survival (P = 0.005). In vitro studies showed that miR-31 promoted ESCC colony formation, migration and invasion. Luciferase reporter and Western blot assays confirmed that three tumour suppressor genes, namely EMP1 (epithelial membrane protein 1), KSR2 (kinase suppressor of ras 2) and RGS4 (regulator of G-protein signalling 4), were targeted by miR-31. We conclude that miR-31 plays oncogenetic functions and can serve as a potential diagnostic and prognostic biomarker for ESCC

Optical Thickness-Encoded Suspension Array for High-Throughput Multiplexed Gene Detection

We proposed a coding and decoding method of suspension array (SA) based on micro-quartz pieces (MQPs) with different optical thicknesses. The capture probes (cDNA) were grafted onto the surfaces of MQPs and specifically recognized and combined with the partial sequence of the target DNA (tDNA) to form a MQP-cDNA-tDNA complex. Quantum dot-labeled signal probes were then used to specifically recognize and bind another portion of the tDNA in the complex to form a double-probe sandwich structure. This optical thickness-encoded SA can be decoded and detected by a dual-wavelength digital holographic phase fluorescence microscope system. We conducted a series of DNA molecule detection experiments by using this encoding method. Control experiments confirmed the specificity of optical thickness-encoded SA in DNA detection. The concentration gradient experiments then demonstrated the response of the MQPs based SA to analyte concentration. Finally, we used the encoding method to detect three types of DNA in a single sample and confirmed the feasibility of the proposed optical thickness-encoded SA in multiplexed DNA detection. The detection results are stable, and the detection exhibits high specificity and good repeatability

In vivo monitoring of microneedle-based transdermal drug delivery of insulin

Soluble microneedles (MNs) have recently become an efficient and minimally invasive tool in transdermal drug delivery because of their excellent biocompatibility and rapid dissolution. However, direct monitoring of structural and functional changes of MNs in vivo to estimate the efficiency of insulin delivery is difficult. We monitored the dissolution of MNs to obtain structural imaging of MNs’ changes by using optical coherence tomography (OCT). We also observed the effect of MNs on microvascular conditions with laser speckle contrast imaging (LSCI) and measured the blood perfusion of skin to obtain functional imaging of MNs. We determined the performance of two soluble MN arrays made from polyvinyl alcohol (PVA) and polyvinyl alcohol/polyvinylpyrolidone (PVA/PVP) by calculating the cross-sectional areas of the microchannels in mouse skin as a function of time. Moreover, the change in blood glucose before and after using MNs loaded with insulin was evaluated as an auxiliary means to demonstrate the ability of the soluble MNs to deliver insulin. Results showed that the structural imaging of these MNs could be observed in vivo via OCT in real time and the functional imaging of MNs could be showed using LSCI. OCT and LSCI are potential tools in monitoring MNs structural and functional changes

Dysregulation of peripheral and intratumoral KLRG1+ CD8+ T cells is associated with immune evasion in patients with non-small-cell lung cancer

Objectives: Killer cell lectin like receptor G1 (KLRG1) is identified as a co-inhibitory receptor for NK cells and antigen-experienced T cells. The role of KLRG1 in immune regulation in patients with non-small cell lung cancer (NSCLC) remains poorly understood. Materials and Methods: We measured the proportion and immune function of KLRG1+CD8+ T cells derived from peripheral blood in patients with NSCLC by flow cytometry. Besides, using data from the gene expression profiles and single-cell sequencing, we explored the expression and immune role of KLRG1 in tumor tissues of patients with NSCLC. We further determined the prognostic value of KLRG1 in terms of overall survival (OS) in NSCLC patients. Results: We found that the proportion of KLRG1+CD8+T cells in peripheral blood significantly increased in patients with NSCLC as compared to those with benign pulmonary nodules and healthy donors. Peripheral KLRG1+CD8+T cell proportion was increased in elder subjects compared to that in younger ones, implying an immunosenescence phenotype. Moreover, the KLRG1+CD8+T cell levels were positively correlated with tumor size and TNM stage in the NSCLC cohort. In vitro stimulation experiments demonstrated that the KLRG1+CD8+T cells from peripheral blood expressed higher levels of Granzyme B and perforin than the KLRG1−CD8+ T cells. However, single-cell RNA sequencing data revealed that the KLRG1+CD8+ T cells were less infiltrated in tumor microenvironment and exhibited impaired cytotoxicity. The KLRG1 gene expression levels were significantly lower in tumor tissues than that in normal lung tissues, and were inversely correlated with CDH1 expression levels. Moreover, higher expression of CDH1 in tumor tissues predicted worse overall survival only in patients with KLRG1-high expression, but not in the KLRG1-low subset. Conclusion: This study demonstrates that KLRG1+CD8+T cells were associated with tumor immune evasion in NSCLC and suggests KLRG1 as a potential immunotherapy target

γδTDEs: An Efficient Delivery System for miR-138 with Anti-tumoral and Immunostimulatory Roles on Oral Squamous Cell Carcinoma

In this study, we sought to investigate the potential application of γδ T cell-derived extracellular vesicles (γδTDEs) as drug delivery system (DDS) for miR-138 in the treatment of oral squamous cell carcinoma (OSCC). Our data showed that overexpression of miR-138 in γδ T cells obtained miR-138-rich γδTDEs accompanying increased expansion and cytotoxicity of γδ T cells. γδTDEs inherited the cytotoxic profile of γδ T cells and could efficiently deliver miR-138 to OSCC cells, resulting in synergetic inhibition on OSCC both in vitro and in vivo. The pre-immunization by miR-138-rich γδTDEs inhibited the growth of OSCC tumors in immunocompetent C3H mice, but not in nude mice, suggesting an immunomodulatory role by miR-13-rich γδTDEs. γδTDEs and miR-138 additively increased the proliferation, interferon-γ (IFN-γ) production, and cytotoxicity of CD8+ T cells against OSCC cells. Only delivered by γδTDEs can miR-138 efficiently target programmed cell death 1 (PD-1) and CTLA-4 in CD8+ T cells. We conclude that γδTDEs delivering miR-138 could achieve synergetic therapeutic effects on OSCC, which is benefited from the individual direct anti-tumoral effects on OSCC and immunostimulatory effects on T cells by both γδTDEs and miR-138; γδTDEs could serve as an efficient DDS for microRNAs (miRNAs) in the treatment of cancer. Keywords: extracellular vesicle, γδ T cell, miRNA, drug delivery system, oral squamous cell carcinom

Gene Expression Profiling in Human Lung Development: An Abundant Resource for Lung Adenocarcinoma Prognosis

<div><p>A tumor can be viewed as a special “organ” that undergoes aberrant and poorly regulated organogenesis. Progress in cancer prognosis and therapy might be facilitated by re-examining distinctive processes that operate during normal development, to elucidate the intrinsic features of cancer that are significantly obscured by its heterogeneity. The global gene expression signatures of 44 human lung tissues at four development stages from Asian descent and 69 lung adenocarcinoma (ADC) tissue samples from ethnic Chinese patients were profiled using microarrays. All of the genes were classified into 27 distinct groups based on their expression patterns (named as PTN1 to PTN27) during the developmental process. In lung ADC, genes whose expression levels decreased steadily during lung development (genes in PTN1) generally had their expression reactivated, while those with uniformly increasing expression levels (genes in PTN27) had their expression suppressed. The genes in PTN1 contain many n-gene signatures that are of prognostic value for lung ADC. The prognostic relevance of a 12-gene demonstrator for patient survival was characterized in five cohorts of healthy and ADC patients [ADC_CICAMS (n = 69, p = 0.007), ADC_PNAS (n = 125, p = 0.0063), ADC_GSE13213 (n = 117, p = 0.0027), ADC_GSE8894 (n = 62, p = 0.01), and ADC_NCI (n = 282, p = 0.045)] and in four groups of stage I patients [ADC_CICAMS (n = 22, p = 0.017), ADC_PNAS (n = 76, p = 0.018), ADC_GSE13213 (n = 79, p = 0.02), and ADC_qPCR (n = 62, p = 0.006)]. In conclusion, by comparison of gene expression profiles during human lung developmental process and lung ADC progression, we revealed that the genes with a uniformly decreasing expression pattern during lung development are of enormous prognostic value for lung ADC.</p></div

The most significantly enriched functional categories and GO terms of genes in each vPTN with corresponding enrichment score (ES).

<p>The most significantly enriched functional categories and GO terms of genes in each vPTN with corresponding enrichment score (ES).</p

Morphological and transcriptomic features of human lung during development.

<p>(<b>A</b>–<b>C</b>), (<b>D</b>–<b>F</b>), (<b>G</b>–<b>I</b>) & (<b>J</b>–<b>L</b>), Morphological images for the four types of human developmental lung samples, i.e., WholeE, EarlyL, MiddleL & MatureL. (<b>M</b>) Cladogram was created with the whole expression profiles obtained for the developmental lung samples and shows the phylogenetic relationships among the developmental lung samples. (<b>N</b>) Hierarchical clustering analysis of top 4000 most divergent genes. For each gene, we calculated its coefficient of variation (CV) based on its expression values across all developmental samples. The genes were then ranked based on their CV values. The heatmap was generated by hierarchical clustering of the top 4000 genes with largest CV values. The colored matrix indicated the relative expression levels of genes (red for higher expression, green for lower). The distribution of samples from each developmental stage was shown above the heatmap. (<b>O</b>) Developmental samples were projected onto the two-dimensional space captured by PCA with the stages of each sample indicated by color.</p