MiR-140-3p inhibits natural killer cytotoxicity to human ovarian cancer via targeting MAPK1

Natural killer (NK) cells have pivotal role in immunotherapy of human ovarian cancer (OC). AlthoughmicroRNAs (miRNAs) participate in dysfunction of NK cells, how and whether miR-140-3p regulates cytotoxicity of NK cells in OC are uncertain. miR-140-3p and mitogen activated protein kinase 1 (MAPK1)abundances were examined via quantitative real-time polymerase chain reaction or western blot. Tumornecrosis factor-a (TNF-a) and interferon-c (IFN-c) abundances were examined via enzyme linkedimmunosorbent assay. NK cytotoxicity to OC was evaluated via lactate dehydrogenase release. The relevanceof miR-140-3p and MAPK1 was proved via luciferase activity analysis. Murine xenograft experiment wasapplied to assess the function of miR-140-3p on NK cytotoxicity. miR-140-3p was elevated and MAPK1 wasdeclined in NK cells from OC patients, while the levels were reversed after treatment of interleukin-2 (IL-2).MiR-140-3p addition mitigated IFN-c and TNF-a production induced via IL-2 as well as NK-92 cytotoxicityto OC cells. Additionally, MAPK1 was negatively regulated via miR-140-3p and ablated the influence of miR140-3p on cytotoxicity, cytokines levels. Besides, miR-140-3p enrichment facilitated tumor growth via suppressing function of NK cells in a xenograft model. miR-140-3p suppressed NK cytotoxicity to OC cells viamediating MAPK1, indicating a new avenue of ameliorating NK cells function for OC treatment.

miR-140-3p inhibits progression of non-small cell lung cancer by targeting Janus kinase 1

microRNAs (miRNAs) have gained more attention due to the biological functions in many cancers, includingnon-small cell lung cancer (NSCLC). However, the roles and the mechanism of miR-140-3p in NSCLCprogression remain poorly understood. In this study, the expression levels of miR-140-3p and Janus kinase 1(JAK1) were measured in NSCLC tissues and cells by quantitative real-time PCR. Cell viability, apoptosis,migration and invasion were detected by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl-trtrazolium bromide, flowcytometry, Western blot or trans-well assay, respectively. Murine xenograft model was conducted to analyzethe anti-tumor effect of miR-140-3p in vivo. Interaction between miR-140-3p and JAK1 was probed byluciferase reporter activity and Western blot. We found that miR-140-3p expression was down-regulated andJAK1 expression was increased in NSCLC tissues and cells compared with those in corresponding controls.Moreover, overexpression of miR-140-3p inhibited cell viability, migration and invasion while promoted cellapoptosis in NSCLC cells and suppressed NSCLC xenograft tumor growth in vivo. Besides, JAK1 was provedas a target of miR-140-3p and its restoration reversed miR-140-3p-mediated regulatory effect on progression ofNSCLC. We concluded that miR-140-3p inhibited NSCLC progression by targeting JAK1, providing a novelavenue for treatment of NSCLC.

miR-140-3p Knockdown Suppresses Cell Proliferation and Fibrogenesis in Hepatic Stellate Cells via PTEN-Mediated AKT/mTOR Signaling

PURPOSE: Liver fibrosis is a major cause of morbidity and mortality and the outcome of various chronic liver diseases. Activation of hepatic stellate cells (HSCs) is the key event in liver fibrosis. Studies have confirmed that miR-140-3p plays a potential regulatory effect on HSC activation. However, whether miR-140-3p mediates the liver fibrosis remains unknown. MATERIALS AND METHODS: Expression of miR-140-3p was detected by real-time quantitative PCR (qPCR). Cell proliferation was measured by MTT, while cell apoptosis rate was determined via flow cytometry. Western blot assay was used to detect the expression of cleaved PARP. The fibrogenic effect was evaluated by expression of α-smooth muscle actin and desmin. Functional experiments were performed in transforming growth factor β1 (TGF-β1)-induced HSC-T6 cells with transfection of anti-miR-140-3p and/or siPTEN. Target binding between miR-140-3p and PTEN was predicted by the TargetScan database and identified using luciferase reporter assay and RNA immunoprecipitation. RESULTS: TGF-β1 induced the activation of HSC-T6 cells, and miR-140-3p expression varied according to HSC-T6 cell activation status. Knockdown of miR-140-3p reduced cell proliferation and the expressions of α-SMA and desmin, as well as increased apoptosis, in TGF-β1-induced HSC-T6 cells, which could be blocked by PTEN silencing. Additionally, inactivation of the AKT/mTOR signaling pathway stimulated by miR-140-3p knockdown was abolished when silencing PTEN expression. PTEN was negatively regulated by miR-140-3p via direct binding in HSC-T6 cells. CONCLUSION: miR-140-3p is an important mediator in HSC-T6 cell activation, and miR-140-3p knockdown suppresses cell proliferation and fibrogenesis in TGF-β1-induced HSC-T6 cells, indicating that miR-140-3p may be a potential novel molecular target for liver fibrosis.

Expression of miR-140-3p in children with autism spectrum disorders and its correlation with cyto-kines / 中华行为医学与脑科学杂志

Objective To investigate the expression of serum miR-140-3p in children with autism spectrum disorder (ASD) and its correlation with interleukin-4 ( IL-4),interleukin-8 (IL-8) and interleu-kin-17A (IL-17A),and provide evidence for the diagnosis and treatment of ASD. Methods Totally 172 ca-ses of ASD children admitted to Anning Hospital of Hainan Province from January 2015 to June 2018 were selected as case group,80 cases of healthy subjects as control group. ASD children were divided into mild-moderate group (n=116) and severe group (n=56) by ASD Behavior Rating Scale and Child Autism Rating Scale. Real-time fluorescence quantitative polymerase chain reaction ( RT-PCR) and enzyme-linked immu-nosorbent assay ( ELISA) were used to detect the changes of serum levels of miR-140-3p,IL-4,IL-8 and IL-17A in each group. ROC curve was used to analyze the diagnostic value of miR-140-3p,IL-4,IL-8 and IL-17A for ASD. Pearson correlation was used to analyze the correlation between serum miR-140-3p and IL-4,IL-8 and IL-17A in children with ASD. Results The serum levels of miR-140-3p ( 0. 86 ± 0. 17 vs 0. 15±0. 03),IL-4(48. 65±13. 82)pg/ml vs (31. 42±7. 50)pg/ml),IL-8((7. 14±2. 05) pg/ml vs (2. 30± 0. 74)pg/ml) and IL-17A((112. 35±51. 64)pg/ml vs (58. 20±26. 40)pg/ml) in the case group were sig-nificantly higher than those in the control group ( P<0. 05). Serum levels of miR-140-3p( 1. 08 ± 0. 24 vs 0. 71±0. 12),IL-4((53. 28±16. 30)pg/ml vs (43. 70±13. 52)pg/ml),IL-8((9. 42±2. 85)pg/ml vs (5. 63 ±1. 48)pg/ml) and IL-17A((138. 40±65. 72)pg/ml vs (96. 74±40. 83) pg/ml) in severe group were sig-nificantly higher than those in mild-moderate group(P<0. 05). The AUC (95%CI) of serum miR-140-3p for ASD diagnosis was 0. 827 (0. 773-0. 886),which were significantly higher than that of IL-4 (0. 719 (0. 651-0. 764)pg/ml),IL-8 (0. 683 (0. 625-0. 743)pg/ml) and IL-17A(0. 745 (0. 683-0. 817)pg/ml). The best cut-off value was 0. 75,and the sensitivity and specificity were 84. 8% and 78. 2%,respectively. The best cut-off value of serum miR-140-3p for diagnosing ASD was 0. 75, and its sensitivity and specificity were 84. 8% and 78. 2%,respectively. The correlation analysis showed that serum miR-140-3p was positively cor-related with IL-4,IL-8 and IL-17A in children with ASD (r=0. 681,r=0. 624,r=0. 775,all P<0. 01). Con-clusion Serum levels of miR-140-3p was significantly higher in ASD children, correlated with levels of IL-4,IL-8 and IL-17A,and miR-140-3p was a potential biomarkers for ASD diagnosis.