Decrease of miR-195 Promotes Chondrocytes Proliferation and Maintenance of Chondrogenic Phenotype via Targeting FGF-18 Pathway

Abstract

Slow growth and rapid loss of chondrogenic phenotypes are the major problems affecting chronic cartilage lesions. The role of microRNA-195 (miR-195) and its detailed working mechanism in the fore-mentioned process remains unknown. Fibroblastic growth factor 18 (FGF-18) plays a key role in cartilage homeostasis; whether miR-195 could regulate FGF-18 and its downstream signal pathway in chondrocyte proliferation and maintenance of chondrogenic phenotypes still remains unclear. The present research shows elevated miR-195 but depressed FGF-18 expressed in joint fluid specimens of 20 patients with chronic cartilage lesions and in CH1M and CH3M chondrocytes when compared with that in joint fluid specimens without cartilage lesions and in CH1W and CH2W chondrocytes, respectively. The following loss of function test revealed that downregulation of miR-195 by transfection of miR-195 inhibitors promoted chondrocyte proliferation and expression of a type II collagen α I chain (Col2a1)/aggrecan. Through the online informatics analysis we theoretically predicted that miR-195 could bind to a FGF-18 3′ untranslated region (3′UTR), also, we verified that a miR-195 could regulate the FGF-18 and its downstream pathway. The constructed dual luciferase assay further confirmed that FGF-18 was a direct target of miR-195. The executed anti-sense experiment displayed that miR-195 could regulate chondrocyte proliferation and Col2a1/aggrecan expression via the FGF-18 pathway. Finally, through an in vivo anterior cruciate ligament transection (ACLT) model, downregulation of miR-195 presented a significantly protective effect on chronic cartilage lesions. Evaluating all of the outcomes of the current research revealed that a decrease of miR-195 protected chronic cartilage lesions by promoting chondrocyte proliferation and maintenance of chondrogenic phenotypes via the targeting of the FGF-18 pathway and that the miR-195/FGF-18 axis could be a potential target in the treatment of cartilage lesions.

Long noncoding RNA MALAT1 regulates HDAC4‐mediated proliferation and apoptosis via decoying of miR‐140‐5p in osteosarcoma cells

“…The procedure was carried out as previously described . HOS and 143B cells after different MALAT1, HDAC4, or miR‐140‐5p intervene were seeded in 96‐well plate and incubated for 48 hours.…”

“…The procedure was carried out as previously described. 29 HOS and 143B cells after different MALAT1, HDAC4, or miR-140-5p intervene were seeded in 96-well plate and incubated for 48 hours. The following assays were performed according to the manufacturer's instructions by applying an EDU detection kit (Keygen).…”

The fibroblast growth factor 8 family in the female reproductive tract

“…In human endothelial cells, hsamiR-505 inhibited FGF18 mRNA and protein levels and inhibited the activity of a luciferase reporter containing the 3′UTR of FGF18 (Yang et al 2014) suggesting that this microRNA interacts directly with FGF18 mRNA. Another miRNA that interacts with the 3′UTR of FGF18 is miR-195 (Wang et al 2017). Whether these miRNAs are involved in reproductive physiology or medicine is just being explored, although miR-505 abundance was found to be downregulated in endometrial carcinoma (Chen et al 2016), a tissue in which increased FGF18 mRNA levels have been observed (see section on 'Gynaecological cancers'.…”

Long non‑coding RNA DANCR promotes HMGA2‑mediated invasion in lung adenocarcinoma cells

“…Immunohistochemical (IHC) staining. The IHC procedure was performed as previously described (31). The LAD tissue specimens were processed as follows: 4% paraformaldehyde fixation, paraffin-embedding, sectioned to 4-µm thickness, deparaffinization, rehydration, hydrogen peroxide incubation, antigen retrieval, blocked in 10% goat serum (Bioworld Technology, Inc., St. Louis Park, MN, USA), primary antibody incubation (anti-HMGA2 and anti-GAPDH) at 4˚C overnight, secondary antibody incubation (goat anti-rabbit IgG H&L; Abcam) at 37˚C for 20 min, streptavidin-horseradish peroxidase complex incubation, diaminobenzidine tetrahydroc hloride (MedChemExpress, Monmouth Junction, NJ, USA) staining, hematoxylin (Amresco, LLC, Solon, OH, USA) and counterstaining.…”