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LncRNA-PCAT1 targeting miR-145-5p promotes TLR4-associated osteogenic differentiation of adipose-derived stem cells.

ABSTRACT: This study was aimed to explore the differential expression of long noncoding RNAs (lncRNA)-PCAT1, miR-145-5p and TLR4 in osteogenic differentiation via the Toll-like receptor (TLR) signalling pathway and consequently determine the potential molecular mechanism. The mRNAs and pathways related to the osteogenic differentiation in human adipose-derived stem cells (hADSCs) were analysed by bioinformatics. The MiRanda and TargetScan database were employed to detect the potential binding sites of miRNAs on lncRNAs and mRNAs. The differential expression of lncRNA-PCAT1, miR-145-5p and TLR4 were detected by qRT-PCR. Rrelated protein expression was analysed by Western blot. The targeted relationships between lncRNA-PCAT1, miR-145-5p and TLR4 were verified by dual-luciferase reporter assay. Alkaline phosphatase (ALP) activity and ARS staining assays were used to measure the impacts exerted by lncRNA PCAT1, miR-145-5p and TLR4 mRNA on osteogenic differentiation. After the induction of osteoblast differentiation, the expression of lncRNA-PCAT1 and TLR4 increased, while the expression of miR-145-5p decreased. Dual-luciferase reporter assay confirmed the targeted relationship between lncRNA-PCAT1, miR-145-5p, and TLR4. LncRNA-PCAT1 negatively regulated miR-145-5p and positively regulated TLR4. Knockdown of lncRNA-PCAT1 or TLR4 decreased the expression of osteogenic differentiation-related proteins, reduced the ALP and ARS levels and the activity of the TLR signalling pathway. MiR-145-5p could reverse the effects of PCAT1 and TLR4 in hADSCs osteogenic differentiation. LncRNA-PCAT1 negatively regulated miR-145-5p, which promoted TLR4 expression to promote osteogenic differentiation by activating the TLR signalling pathway.

SUBMITTER: Yu L

PROVIDER: S-EPMC6237555 | biostudies-literature | 2018 Dec

REPOSITORIES: biostudies-literature

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LncRNA-PCAT1 targeting miR-145-5p promotes TLR4-associated osteogenic differentiation of adipose-derived stem cells.

Yu Lingjia L Qu Hao H Yu Yifeng Y Li Wenjing W Zhao Yu Y Zhao Yu Y Qiu Guixing G

Journal of cellular and molecular medicine 20181019 12

This study was aimed to explore the differential expression of long noncoding RNAs (lncRNA)-PCAT1, miR-145-5p and TLR4 in osteogenic differentiation via the Toll-like receptor (TLR) signalling pathway and consequently determine the potential molecular mechanism. The mRNAs and pathways related to the osteogenic differentiation in human adipose-derived stem cells (hADSCs) were analysed by bioinformatics. The MiRanda and TargetScan database were employed to detect the potential binding sites of miR ...[more]

PMID: 30338912

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Project description:BackgroundLong-non-coding RNA PVT1 (lncRNA PVT1) can be used as an oncogenic regulatory non-coding RNA (ncRNA) for many cancers. However, its function and mechanism in breast cancer (BRCA) are still not clearly elucidated.ObjectiveWe attempt to explain the mechanism of PVT1's role in breast cancer from different perspectives.MethodsWe analyzed the expression of PVT1 and its correlation with the breast cancer related clinical data in the The Cancer Genome Atlas (TCGA) database. We used PVT1 overexpression and knockdown lentivirus to infect breast cancer MDA-MB-231 cell line for cell function verification, in vitro using CCK-8 to measure proliferation, flow cytometry to measure apoptosis, transwell test to measure invasion and migration ability, detecting cell extracellular acidification rate (ECAR) to assess glycolysis metabolism and explore the biological functions of PVT1 in breast cancer cells. Transcriptome sequencing was used to analyze the changes of related genes in cells after overexpression of PVT1. In vivo we used a xenograft model to study the effect of PVT1 on breast cancer.ResultsPVT1 was up-regulated in breast cancer tissues and was positively correlated with the clinical stage of breast cancer patients. Overexpression of PVT1 in vitro promoted cell proliferation, migration and invasion, and promoted tumor growth in vivo. Knockdown of PVT1 led to the opposite biological consequence. Further bioinformatics analysis showed that PVT1 changes the glycolysis metabolism of tumors through regulation of glycolysis-related genes. In addition, the expression of miR-145-5p is negatively correlated with PVT1. We consider the possibility of PVT1 promoting cell proliferation and metastasis by regulating the aerobic glucose metabolism in breast cancer cells through sponging the miR-145-5p.ConclusionOur results reveal a potential pathway for competing endogenous RNA to regulate breast cancer glucose metabolism. PVT1 regulates glycolysis related genes expression by competitively binding to endogenous miR-145-5p in breast cancer cells to change the metabolic phenotype. This may Provide new ideas for precise molecular therapy targets for breast cancer.

Dataset Information

LncRNA-PCAT1 targeting miR-145-5p promotes TLR4-associated osteogenic differentiation of adipose-derived stem cells.

Publications

LncRNA-PCAT1 targeting miR-145-5p promotes TLR4-associated osteogenic differentiation of adipose-derived stem cells.

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