Whole transcriptome sequencing analysis of blood plasma-derived exosomes from immune-related hearing loss.

BACKGROUND: Early diagnosis of immune-related hearing loss and timely treatment can prevent structural damage to the inner ear and contribute to hearing retention. Exosomal miRNAs, lncRNAs and proteins have great prospects as novel biomarkers for clinical diagnosis. Our study aimed to investigate the molecular mechanisms of exosomes or exosomal ceRNA regulatory networks in immune-related hearing loss. METHODS: An immune-related hearing loss mice model was constructed by injection with inner ear antigen, and then the blood plasma samples of the mice were collected for exosomes isolation by ultra-centrifugation. Subsequently, the different exosomes were sent for whole transcriptome sequencing using Illumina platform. Finally, a ceRNA pair was chosen for validation by RT-qPCR and dual luciferase reporter gene assay. RESULTS: The exosomes were successfully extracted from the blood samples of the control and the immune-related hearing loss mice. After sequencing, 94 differentially expressed (DE) lncRNAs, 612 DEmRNAs, and 100 DEmiRNAs were found in the immune-related hearing loss-associated exosomes. Afterwards, ceRNA regulatory networks consisting of 74 lncRNAs, 28 miRNAs and 256 mRNAs were proposed, and the genes in the ceRNA regulatory networks were significantly enriched in 34 GO terms of biological processes and 9 KEGG pathways. Finally, Gm9866 and Dusp7 were significantly up-regulated, while miR-185-5p level was declined in the exosomes from immune-related hearing loss, and Gm9866, miR-185-5p and Dusp7 interacted with each other. CONCLUSIONS: Gm9866-miR-185-5p-Dusp7 was confirmed to be closely correlated with the occurrence and progression of immune-related hearing loss.

Visible-Light-Induced Difluoroalkylation of Alkenes and Alkynes with Fluoro-Containing Hypervalent Iodane (III) Reagents Under Photo-Catalyst-Free Conditions.

A visible-light-induced difluoroalkylation of unactivated alkenes by fluoro-containing hypervalent iodine-based difluoroalkylation reagent was achieved for the first time under photo-catalyst-free conditions. Moreover, the same reaction conditions were applicable to the difluoroalkylation of alkynes to give the hydrodifluoroalkylation products in moderate to excellent yields. The readily available reagent, broad substrate scope, and photo-catalyst-free conditions make this protocol an efficient and environmental friendly method for the hydrodifluoroalkylation of alkenes and alkynes.

Photoinduced Three-Component Difluoroalkylation of Quinoxalinones with Alkenes via Difluoroiodane(III) Reagents.

An environmentally friendly strategy for the photocatalyzed three-component reaction between quinoxalinones, alkenes, and hypervalent iodine(III) reagents is disclosed. The new designed difluoroiodane(III) reagent shows excellent reactivity, providing a wide range of difluoroalkyl-substituted quinoxaline-2(1H)-ones in moderate to excellent yields under mild conditions. Experimental studies demonstrated that a difluoroalkyl radical intermediate was involved in this reaction.

miR-183/TMSB4Y, a new potential signaling axis, involving in the progression of laryngeal cancer via modulating cell adhesion.

Laryngeal cancer (LCa) is a prevalent malignant head and neck cancer with relatively unclear pathogenesis. A prior study has suggested that miR-183 differentially expressed in laryngeal-related malignancies, but its accurate role has not been fully ascertained in LCa. miR-183 expression in LCa tissues and cells was detected assisted by TCGA/GEO databases or qRT-PCR assay, relatively. Target genes of miR-183 were predicted via accessing to TargetScan website. Luciferase activity analysis was conducted to determine the relationship between miR-183 and its possible target. CCK-8, colony formation and transwell invasion and migration experiments were implemented to measure LCa cell viability, invasion and migration. Western blot assay was utilized to evaluate cell adhesion and EMT-related proteins expressions. The expression of miR-183 was expressed in LCa tissue samples and cells at higher levels than normal controls. Upregulation of miR-183 facilitated Hep-2 and TU212 cells viability, while miR-183 reduction inhibited the proliferative potential of Hep-2 and TU212 cells. TMSB4Y was determined as a possible target of miR-183, and its expression was decreased in LCa. LCa patients with low TMSB4Y expression had poorer outcomes relative to that with high TMSB4Y expression. TMSB4Y overturned the promoting impacts of miR-183 on the LCa cellular malignant behaviors, including cell proliferation, colonogenicity, invasion and migration. miR-183 overexpression inhibited cell adhesion through inhibiting TMSB4Y expression. Overall, all results elucidated that miR-183, as an oncogenic molecule in LCa, may be used to predict the prognosis of LCa patients by targeting TMSB4Y.

LncRNA LINC00888 upregulation predicts a worse survival of laryngeal cancer patients and accelerates the growth and mobility of laryngeal cancer cells through regulation of miR-378g/TFRC.

In this study, we aimed to detect the clinical significance and potential mechanism of LINC00888 in laryngeal cancer. Data from The Cancer Genome Atlas (TCGA) afforded the expression of LINC00888 in laryngeal cancer samples. The clinical significance of LINC00888 expression in laryngeal cancer was demonstrated by χ2 , Cox analysis, and Kaplan-Meier. The downstream targets of LINC00888 were identified based on analysis from bioinformatics and laryngeal cancer-related TCGA data sets. The microRNA-378g (miR-378g)/TFRC (transferrin receptor) axis was selected and identified by qRT-PCR, Western blot analysis, and luciferase activity assays. Cell counting kit-8, colony formation, and transwell assays were applied to detect the phenotypes of laryngeal cancer cells. We observed that LINC00888 expression was notably increased in laryngeal cancer and associated with death, recurrence, and prognosis. Depletion of LINC00888 repressed the proliferative and motile abilities of laryngeal cancer cells in vitro. LINC00888 was predicted to act as a competing endogenous RNA within the microRNA (miRNA)/messenger RNA (mRNA) axis based on analysis from bioinformatics and laryngeal cancer-related TCGA data sets. Interestingly, we discovered that LINC00888 functioned as an miRNA sponge to suppress the effect of miR-378g on laryngeal cancer cells behaviors, as well as positively regulate TFRC expression. Furthermore, the knockdown of TFRC strengthened the inhibitory effect of si-LINC00888 on laryngeal cancer cells' malignant properties. LINC00888 is an oncogenic lncRNA that promotes the growth and mobility of laryngeal cancer cells by controlling laryngeal cancer-related mRNA and tumor-suppressive miRNA. The LINC00888/miR-378g/TFRC pathway might lead to the development of laryngeal cancer cells and, therefore, might be a candidate therapeutic target for laryngeal cancer.