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The role of microRNA-3085 in chondrocyte function
Get PDFMicroRNAs have been shown to play a role in cartilage development, homeostasis and breakdown during osteoarthritis. We previously identified miR-3085 in humans as a chondrocyte-selective microRNA, however it could not be detected by Northern blot. The aim of the current study was to prove that miR-3085 is a microRNA and to investigate the function of miR-3085 in signaling pathways relevant to cartilage homeostasis and osteoarthritis. Here, we confirm that miR-3085 is a microRNA and not another class of small RNA using (1) a pre-miR hairpin maturation assay, (2) expression levels in a Dicer null cell line, and (3) Ago2 pulldown. MicroRNA-3085-3p is expressed more highly in micromass than monolayer cultured chondrocytes. Transfection of miR-3085-3p into chondrocytes decreases expression of COL2A1 and ACAN, both of which are validated as direct targets of miR-3085-3p. Interleukin-1 induces the expression of miR-3085-3p, at least in part via NFκB. In a feed-forward mechanism, miR-3085-3p then potentiates NFκB signaling. However, at early time points after transfection, its action appears to be inhibitory. MyD88 has been shown to be a direct target of miR-3085-3p and may be responsible for the early inhibition of NFκB signaling. However, at later time points, MyD88 knockdown remains inhibitory and so other functions of miR-3085-3p are clearly dominant. TGFβ1 also induces the expression of miR-3085-3p, but in this instance, it exerts a feedback inhibition on signaling with SMAD3 and SMAD4 shown to be direct targets. This in vitro analysis shows that miR-3085-3p functions in chondrocytes to induce IL-1-signaling, reduce TGFβ1 signaling, and inhibit expression of matrix genes. These data suggest that miR-3085-3p has a role in chondrocyte function and could contribute to the process of osteoarthritis
The role of microRNA-3085 in chondrocyte function
Get PDFMicroRNAs have been shown to play a role in cartilage development, homeostasis and breakdown during osteoarthritis. We previously identified miR-3085 in humans as a chondrocyte-selective microRNA, however it could not be detected by Northern blot. The aim of the current study was to prove that miR-3085 is a microRNA and to investigate the function of miR-3085 in signaling pathways relevant to cartilage homeostasis and osteoarthritis. Here, we confirm that miR-3085 is a microRNA and not another class of small RNA using (1) a pre-miR hairpin maturation assay, (2) expression levels in a Dicer null cell line, and (3) Ago2 pulldown. MicroRNA-3085-3p is expressed more highly in micromass than monolayer cultured chondrocytes. Transfection of miR-3085-3p into chondrocytes decreases expression of COL2A1 and ACAN, both of which are validated as direct targets of miR-3085-3p. Interleukin-1 induces the expression of miR-3085-3p, at least in part via NFκB. In a feed-forward mechanism, miR-3085-3p then potentiates NFκB signaling. However, at early time points after transfection, its action appears to be inhibitory. MyD88 has been shown to be a direct target of miR-3085-3p and may be responsible for the early inhibition of NFκB signaling. However, at later time points, MyD88 knockdown remains inhibitory and so other functions of miR-3085-3p are clearly dominant. TGFβ1 also induces the expression of miR-3085-3p, but in this instance, it exerts a feedback inhibition on signaling with SMAD3 and SMAD4 shown to be direct targets. This in vitro analysis shows that miR-3085-3p functions in chondrocytes to induce IL-1-signaling, reduce TGFβ1 signaling, and inhibit expression of matrix genes. These data suggest that miR-3085-3p has a role in chondrocyte function and could contribute to the process of osteoarthritis
Roles of miR-3085-3p in skeletal development and osteoarthritis
Get PDFOsteoarthritis (OA) is a progressive arthritis catheterized by cartilage degradation, subchondral bone sclerosis and synovial inflammation. MicroRNAs are endogenous non-coding short RNAs, which repress gene expression. Numerous microRNAs have been reported to be involved in pathogenesis of OA. MiR-3085-3p is a novel microRNA that is selectively expressed in human cartilage and targets integrin α5 (ITGA5). This project ravelled roles of miR-3085-3p in the skeletal development and progression of OA, following the optimization of time and dosage across experiments.
Overexpression of miR-3085-3p decreased expression of ITGA5 in DF1, SW1353, HeLa and C28/I2 in various assays including qRT-PCR, western blotting and luciferase assay. In luciferase assay, 50nM miR-3085-3p mimic and 72h transfection time are optimal. In qRT-PCR, 50nM and a 48h is recommended for either SW1353 or HeLa, whilst 100nM for C28/I2. In western blotting, 100nM and 72h are reliable, where C28/I2 is not applicable.
MiR-3085-3p regulated cartilage homeostasis and chondrocyte function through decreasing the expression of ACAN and COL2A1, and various signalling pathways. MiR-3085-3p increased IL-1β induced MMP13, but transiently downregulated NFκB signalling by targeting MYD88. It repressed TGFβ/SMAD signalling by targeting SMAD2, SAMD3, and SMAD4. MiR-3085-3p potentiates WNT3A-induced TOPFLASH reporter as well as WNT3A-induced AXIN2.
MiR-3085-3p has roles in hMSC differentiation. It could depress chondrogenesis and promote IL-1 induced MMP-13 via JNK/AP-1 pathway in primary HACs. MiR-3085-3p also negatively regulates adipogenesis by inhibiting adipogenic marker genes - CEBPα and PPARγ. In contrast, miR-3085-3p enhances osteogenesis by directly targeting CMTM3 and LBH, especially CTDSP2
We did not successfully generate the miR-3085-3p null mice, as a consequence of embryonic lethality. Lethality was also observed in the ITGA5 cartilage-specific knockout mice. Significant developmental defects in skeletal development and possible accelerated progression of osteoarthritis were revelled in ITGA5 cartilage-specific knockout mice, suggesting that ITGA5 is crucial in these processes. MiR-3085-3p potentially has a negative influence on skeletal development and osteoarthritis by targeting ITGA5. defects in skeletal development and possible accelerated progression of osteoarthritis were revelled in ITGA5 cartilage-specific knockout mice, suggesting that ITGA5 is crucial in these processes. MiR-3085-3p potentially has negative effects on skeletal development and osteoarthritis by targeting ITGA5
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Cansó nova, curiosa y divertida [Texto impreso] : ab estrivillo, pera cantar lo Joven del dia
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Chiste de les Filaneres [Texto impreso]
Get PDFHay un ejemplar encuadernado con: Romans, y coloqui nou, pera divertir el humor y desterrar la melancolia, yà que no tenim dinès ... (NP849.91/3085).Contiene : Trovo
Segon rahonament que tingueren Don Pascasio y Patatorta [Texto impreso] : abentse tornat à encontrar una vesprada en la alameda, y no podentse negar Patatorta al argument, comensà Don Pascasio de este modo
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Chiste nou y divertit [Texto impreso] : Escoltenme señores qu'els apresisa y oirán el gran chasco que es cas de risa. Este ha pasat en un poble qu'es molt nomenat, y Alboraya es diu.
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Título: y Roquét y Goriét Mechorals en les festes dels Sants de la Pedra,.
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