Dataset Information

Exosomes from LNCaP cells promote osteoblast activity through miR-375 transfer.

ABSTRACT: Previous studies have revealed that exosomes influence tumour metastasis, diagnosis and treatment. In addition, exosomal microRNAs (miRNAs/miRs) are closely associated with the metastatic microenvironment; however, the regulatory role of exosomal miRNAs from prostate cancer cells on bone metastasis remains poorly understood. In the present study, a series of experiments were performed to determine whether exosomal miR-375 from LNCaP cells promote osteoblast activity. Exosomes were isolated and purified by ultracentrifugation, total RNA from cells and total miRNA from exosomes were then extracted, and miR-375 levels were detected by reverse transcription-quantitative polymerase chain reaction. Exosome libraries from LNCaP and RWPE-1 cells were sequenced and selected using an Illumina HiSeq™ 2500 system. The effects of exosomes on osteoblasts were determined and osteoblast activity was evaluated by measuring the activity of alkaline phosphatase, the extent of extracellular matrix mineralisation and the expression of osteoblast activity-associated marker genes. Morphological observations, particle size analysis and molecular phenotyping confirmed that cell supernatants contained exosomes. Differential expression analysis confirmed high miR-375 expression levels in LNCaP cell-derived exosomes. The ability of exosomes to enter osteoblasts and increase their levels of miR-375 was further analysed. The results demonstrated that exosomal miR-375 significantly promoted osteoblast activity. In conclusion, the present study may lead to further investigation of the function role of exosomal miR-375 in the activation and differentiation of osteoblasts in PCa.

SUBMITTER: Li SL

PROVIDER: S-EPMC6447935 | biostudies-literature | 2019 May

REPOSITORIES: biostudies-literature

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Exosomes from LNCaP cells promote osteoblast activity through miR-375 transfer.

Li Su-Liang SL An Na N Liu Bing B Wang Sheng-Yu SY Wang Jian-Jun JJ Ye Yun Y

Oncology letters 20190305 5

Previous studies have revealed that exosomes influence tumour metastasis, diagnosis and treatment. In addition, exosomal microRNAs (miRNAs/miRs) are closely associated with the metastatic microenvironment; however, the regulatory role of exosomal miRNAs from prostate cancer cells on bone metastasis remains poorly understood. In the present study, a series of experiments were performed to determine whether exosomal miR-375 from LNCaP cells promote osteoblast activity. Exosomes were isolated and p ...[more]

PMID: 30988815

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Project description:BackgroundTendon derived stem cells (TDSCs) have proven to be effective in tendon repair by secreting paracrine factors, which modulate the function of resident cells and inflammatory process. Exosomes, which are secreted from cells to mediate intercellular communication, may be used to treat tendon injuries. Here, we aimed to determine the effects of exosomes from TDSCs (TDSC-Exos) on tendon repair and to explore the underlying mechanism by investigating the role of microRNAs (miRNAs).MethodsTDSC-Exos were isolated from TDSC conditioned medium. In vitro studies were performed to investigate the effects of TDSC-Exos on the proliferation, migration, cytoprotection, collagen production and tendon-specific markers expression in tenocytes. In order to determine the therapeutic effects of TDSC-Exos in vivo, we used a scaffold of photopolymerizable hyaluronic acid (p-HA) loaded with TDSC-Exos (pHA-TDSC-Exos) to treat tendon defects in the rat model. Subsequently, RNA sequencing and bioinformatic analyses were used to screen for enriched miRNAs in TDSC-Exos and predict target genes. The miRNA-target transcript interaction was confirmed by a dual-luciferase reporter assay system. In order to determine the role of candidate miRNA and its target gene in TDSC-Exos-regulated tendon repair, miRNA mimic and inhibitor were transfected into tenocytes to evaluate cell proliferation and migration.ResultsTreatment with TDSC-Exos promoted proliferation, migration, type I collagen production and tendon-specific markers expression in tenocytes, and also protected tenocytes from oxidative stress and serum deprivation. The scaffold of pHA-TDSC-Exos could sever as a sustained release system to treat the rat model of tendon defects. In vivo study showed that TDSC-Exos promoted early healing of injured tendons. Rats treated with TDSC-Exos had better fiber arrangement and histological scores at the injury site. Besides, the injured tendons treated with TDSC-Exos had better performance in the biomechanical testing. Therefore, the pHA-TDSC-Exos scaffold proved to facilitate tendon repair in the rat model. miR-144-3p was enriched in TDSC-Exos and promoted tenocyte proliferation and migration via targeting AT-rich interactive domain 1A (ARID1A).ConclusionsTDSC-Exos enhanced tenon repair through miR-144-3p-regulated tenocyte proliferation and migration. These results suggest that TDSC-Exos can serve as a promising strategy to treat tendon injuries.

Dataset Information

Exosomes from LNCaP cells promote osteoblast activity through miR-375 transfer.

Publications

Exosomes from LNCaP cells promote osteoblast activity through miR-375 transfer.

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