MiRNA-145 suppresses lung adenocarcinoma cell invasion and migration by targeting N-cadherin.

OBJECTIVE: To investigate the roles of miR-145 in lung adenocarcinoma (LAC) and to clarify the regulation of N-cadherin by miR-145. RESULTS: In 57 paired clinical LAC tissues, diminished miR-145 was significantly correlated with the lymph node metastasis and was negatively correlated with N-cadherin mRNA level expression. Wound healing and transwell assays revealed a reduced capability of tumor metastasis induced by miR-145 in LAC. miR-145 negatively regulated the invasion of cell lines through targeting N-cadherin by directly binding to its 3'-untranslated region. Silencing of N-cadherin inhibited invasion and migration of LAC cell lines similar to miR-145 overexpression. CONCLUSIONS: MiR-145 could inhibit invasion and migration of lung adenocarcinoma cell lines by directly targeting N-cadherin.

Association Between Serum Trace Heavy Metals and Liver Function Among Adolescents.

BACKGROUND: Exposure to metals has been associated with liver-related disease. Few studies have explored the effect of sex stratification on adolescent liver function. METHOD: From the National Health and Nutrition Examination Survey (2011-2016), 1143 subjects aged 12-19 years were selected for analysis. The outcome variables were the levels of alanine aminotransferase (ALT), aspartate aminotransferase, and gamma-glutamyl transpeptidase. RESULTS: The results showed a positive association between serum zinc and ALT in boys (odds ratio [OR], 2.37; 95% confidence interval [CI], 1.11-5.06). Serum mercury was associated with an increase in ALT level in girls (OR, 2.73; 95% CI, 1.14-6.57). Mechanistically, the efficacy mediated by total cholesterol accounted for 24.38% and 6.19% of the association between serum zinc and ALT. CONCLUSIONS: The results imply that serum heavy metals were associated with the risk of liver injury, possibly mediated by serum cholesterol, in adolescents.

Association between mixed exposure of polycyclic aromatic hydrocarbons and telomere length in general population: NHANES 2001-2002.

Although an association between single polycyclic aromatic hydrocarbons (PAHs) adult exposure and telomere length has been reported, the evidence of mixed PAHs (1-napthol, 2-napthol, 3-fluorene, 2-fluorene, 3-phenanthrene, 1-phenanthrene, 2-phenanthrene, and 1-pyrene) exposure and telomere length in the adult general population is still not clear. A total of 1460 adults over the age of 20 years provided urine information on 8 PAHs and selected covariates from the 2001-2002 National Health and Nutrition Examination Survey (NHANES). Bayesian nuclear machine regression (BKMR) was conducted to analyze these associations of telomere length in multiple PAH-exposed environments. Linear regression is mainly used for correlation analysis of PAHs with selected covariate adjustments. Restricted cubic spline (RCS) is used to estimate the correlation between selected PAHs and telomere length. After adjusting for potential covariates, PAHs mixed exposure was negatively associated with telomere length. The linear regression results showed that 2-napthol and 2-fluorene were negatively correlated with telomere length. Telomere length decreased by 1.0% in the fully adjusted model per increment of one unit in the base-10-logarithm-transformed 2-napthol and 2-fluorene concentrations (P = 0.030 and 0.049, respectively). However, the other 6 PAH metabolites were not significantly different. In addition, RCS results showed that 2-napthol has a marginal dose effect relationship with telomere length. Our present study suggested that PAHs are negatively associated with telomere length in the general population of the USA. Considering that the low level of PAHs exposure in the general population can also induce reduced telomere length and potential health risks, future research is needed to explore potential mechanisms.

[The decreased proportion of CD4+CD25+FOXP3+ regulatory T cells in peripheral blood of children with juvenile idiopathic arthritis and its negative correlation with cytokine level].

Objective To investigate the expression of CD4+CD25+FOXP3+regulatory T cells(Tregs), IL-6 and TNF-α in patients with juvenile idiopathic arthritis (JIA) and its clinical significance. Methods The percentages of CD4+CD25+FOXP3+ Tregs in 58 children with JIA and 40 healthy controls were detected by flow cytometry. Serum levels of IL-6 and TNF-α in each patient was detected by chemiluminescence. The correlations between the expression of CD4+CD25+FOXP3+ Tregs and IL-6, TNF-α were analyzed by pearson correlation analysis. We measured the expression level of CD4+CD25+FOXP3+regulatory T cells, IL-6 and TNF-α of 18 cases six weeks after the treatment of tocilizumab (TCZ) in order to figure out the dynamic changes using methods above. Results The percentages of CD4+CD25+FOXP3+Tregs in juvenile idiopathic arthritis were significantly lower than those in 40 healthy volunteers, while levels of IL-6 and TNF-α were significantly higher. However, no obvious difference in the levels of CD4+CD25+FOXP3+Tregs, IL-6 or TNF-α was observed between patients with systemic and poly-articular JIA. Pearson correlation analysis showed that the percentages of CD4+CD25+FOXP3+Tregs negatively correlated with the levels of IL-6 and TNF-α, while levels of IL-6 positively correlated with the levels of TNF-α. Compared with pre-treatment of TCZ, levels of CD4+CD25+FOXP3+Tregs in post-treatment markedly increased, which however were still lower than control group while the levels of IL-6 significantly decreased, yet remained higher than control group. There was no statistical difference between post and pre-treatment in the levels of TNF-α. Conclusion The percentages of CD4+CD25+FOXP3+ Tregs in peripheral blood of JIA children decreases, and it has a negative correlation with IL-6 and TNF-α. Furthermore, the levels of CD4+CD25+FOXP3+ Tregs and IL-6 are partially restored after treatment with TCZ, which may be helpful to assess the activity of systemic JIA and the efficacy of therapy.

FLT3LG and IFITM3P6 consolidate T cell activity in the bone marrow microenvironment and are prognostic factors in acute myelocytic leukemia.

Acute myelocytic leukemia (AML) is a malignancy of the stem cell precursors of the myeloid lineage. CD4+ and CD8+ T cells play pivotal roles in influencing AML progression but are functionally suppressed in the bone marrow microenvironment. We aimed to find hub genes related to T cell exhaustion and suppression, thereby providing evidence for immunotherapy. In this study, gene transcriptome expression data from TCGA and TARGET databases were utilized to find key genes. Firstly, CIBERSORT immune cell infiltration algorithm and WGCNA method were used to identify CD4+ and CD8+ T cells-related genes. Univariate and multivariate cox regression analyses were then introduced to construct the overall survival prognosis model and included hub genes. The ESTIMATE and ssGSEA scoring methods were used to analyze the correlation between the hub genes and immune activity. Single-cell transcriptome analysis was applied to detect the immune cells expressing hub genes, hence, to detect exact mechanisms. Consequently, FLT3LG and IFITM3P6 were determined to be positively correlated with patients' overall survival and microenvironment immune activity. Further study suggested FLT3-FLT3LG and IFITM3P6-miR-6748-3p-CBX7 signaling axes were involved in CD4+ and CD8+ T cells activation. This may be one of the mechanisms of T cells suppression in AML.

Neuromedin U and neurotensin may promote the development of the tumour microenvironment in neuroblastoma.

Stage 4S neuroblastoma, as defined by the International Neuroblastoma Staging System committee (INSS), is known to regress spontaneously and have a more favourable outcome compared with stage 4 tumours. Comparing the molecular differences between these two stages may provide insights into the progression of neuroblastoma. Our study aimed to explore the molecular differences in the tumour microenvironment (TME) between INSS stage 4S and stage 4 tumours to provide an insight into the mechanisms underlying the biological processes of neuroblastoma. We downloaded the datasets GSE120572 and GSE73517 from the GEO database and pre-processed them using the limma package. CIBERSORT deconvolution agorithm was applied to analyse the differences in 22 infiltrating immune leukocyte subsets between the two stages. We used gene ontology (GO) enrichment analysis to determine the biological process (BP) annotation of differentially expressed genes (DEGs) using the online WebGestalt tool. Hub genes were determined in the STRING database and Cytoscape, and the expression of these genes was verified in the Oncomine database. Then these critical genes were performed survival analysis in TARGET database. We further validated the hub genes using a transwell assay and wound healing assay to detect the function of the genes in the neuroblastoma cell line SK-N-BE(2). GO analysis revealed that the 216 DEGs between stage 4S and stage 4 were enriched in aggressive biological processes. Neuromedin U (NMU) and neurotensin (NTS), which were significantly associated with patients' overall survival rate, were verified to be elevated in stage 4, and to promote the proliferation and invasion of the SK-N-BE(2) cell. Tumour infiltrating leukocyte analysis showed a high infiltration of regulatory T cells and type 2 tumour-associated macrophages in stage 4 but not in stage 4S. Results of gene co-expression correlation, and the results of previous studies, suggest that NMU and NTS may play certain roles in modulating TME, thus facilitating the progression of neuroblastoma.

In Vitro Methods for Analyzing miRNA Roles in Cancer Cell Proliferation, Invasion, and Metastasis.

MicroRNAs (miRNAs) are small noncoding, single-stranded RNAs consisting of 20-24 nucleotides (Bartel, Cell 116:281-297, 2004), which regulate target genes expression by interacting with 3'-untranslated regions (3'-UTRs) of target mRNAs, leading to translation repression or mRNA degradation (Filipowicz et al., Nat Rev Genet 9:102-114, 2008; Nilsen, Trends Genet 23:243-249, 2007). Accumulating evidence has elucidated them as oncogenes or tumor suppressors in cancers such as hepatocellular carcinoma, breast cancer, and lung cancer (Liu et al., Gastroenterology 136:683-693, 2009; Yu et al., Cell 131:1109-1123, 2007; Zhou et al., Sci Rep 7:42680, 2017; Iorio and Croce, Carcinogenesis 33:1126-1133, 2012). MiRNAs are involved in various biological processes, including cell proliferation (Liu et al., Mol Cancer Res 11:1314-1325, 2013), differentiation (Liu et al., Mol Cancer Res 11:1314-1325, 2013), apoptosis (Pan et al., Oncol Res 24:429-435, 2016), invasion and metastasis (Liu et al., Nat Commun 8:14270, 2017). Understanding the role of miRNAs in tumor gives new perspective on cancer diagnosis and therapy (Rupaimoole and Slack, Nat Rev Drug Discov 16:203-222, 2017; Berindan-Neagoe et al., CA Cancer J Clin 64:311-336, 2014). This chapter will focus on the in vitro methods for identifying miRNAs roles in cell proliferation, invasion, and metastasis in tumor development, which includes CCK-8 assay, Wound Healing assay, and Transwell assay.

miR-1290 promotes lung adenocarcinoma cell proliferation and invasion by targeting SOCS4.

miRNAs play important roles in lung adenocarcioma (LADC) progression. We previously found that miR-1290 expression was upregulated in LADC tissue and serum samples from patients with LADC, and correlated with prognosis. However, the biological role of miR-1290 in LADC and mechanism of such role are poorly understood. Here, we found that miR-1290 overexpression promoted LADC cell proliferation, cell cycle progression and invasion, while suppressing cell apoptosis in vitro. Furthermore, miR-1290 promoted tumor growth, invasion and metastasis in vivo. miR-1290 downregulated suppressor of cytokine signaling 4 (SOCS4) at both the mRNA and protein levels by targeting SOCS4. Reduced SOCS4 level reversed the inhibitory effect of miR-1290 downregulation on cell proliferation and invasion. miR-1290 activated the JAK/STAT3 and PI3K/AKT signaling pathways by targeting SOCS4. An inverse correlation was observed between miR-1290 and SOCS4 expression in LADC tissues. Clinicopathological characteristics analysis showed that SOCS4 expression was negatively associated with higher clinical stage and lymph node metastasis. These observations suggest that miR-1290 promotes LADC cell proliferation and invasion by targeting SOCS4.