MiR-1297 Inhibits Proliferation And Tumor Growth In Osteosarcoma By Targeting CCND2 And Efficacy And Safety Of Mesenchymal Stem Cell Injection For Patients With Osteoarthritis:A Meta-analysis

Objective: To investigate the effect of miR-1297 on the proliferation and cycle of cells in vitro and tumor growth in vivo by targeting CCND2 in osteosarcoma.Methods:(1)The relative expression of miR-1297 in osteosarcoma cells was detected by real-time quantitative polymerase chain reaction(qRT-PCR).(2)The target genes of miR-1297 were predicted by bioinformatics websites miRDB,miBase and miRDIP.(3)The relative expression of CCND2 in osteosarcoma cell lines was determined by qRTPCR and Western blotting.(4)The effect of miR-1297 on the proliferation of osteosarcoma cells was investigated by EdU assay and clone formation after transfection of miR-1297 mimic and miR-1297 inhibitor.(5)Flow cytometry assay was performed to investigate the effect of miR-1297 on the cell cycle of osteosarcoma cells after transfection of miR-1297 mimic and miR-1297 inhibitor.(6)The efficiency of siRNA of CCND2 was verified by qRT-PCR and Western blotting.(7)The effect of CCND2 on the proliferation of osteosarcoma cells was investigated by EdU and clone formation test after knockdown of CCND2 expression.(8)Cell cycle of osteosarcoma was detected by flow cytometry after knockdown of CCND2.(9)Co-transfection of miR-1297 mimic and CCND2 overexpression plasmid was conducted to demonstrate that the inhibition of miR-1297 could be rescued by CCND2.(10)The cells stably-overexpress of miR-1297 were constructed to explore the effect of miR-1297 on the growth of osteosarcoma in vivo.Results: The qRT-PCR detection showed that miR-1297 was lowly expressed in four osteosarcoma cells.The up-regulation of miR-1297 could significantly inhibit the proliferation of osteosarcoma cells and arrest the tumor cell cycle in the G1 phase,while its inhibitors showed the opposite effect.The expression of mRNA and protein showed that CCND2 was significantly upregulated in the three osteosarcoma cells.Knocking down the expression of CCND2 in tumor cells could significantly impede the proliferation of cells and block most cells in the G1 phase.Luciferase reporter assay demonstrated that miR-1297 could bind to the 3 ’UTR of CCND2.The inhibition of miR-1297 could be rescued by CCND2 overexpressing plasmid in osteosarcoma cells.It was found that HOS cells with stable expression of miR-1297 were constructed by lentivirus to form tumor in subcutaneous and tibia respectively in vivo.The results showed that the tumor size and weight of miR-1297 group were significantly lower than the control group.Conclusion: miR-1297 could inhibit the proliferation of osteosarcoma cells and tumor growth in vivo by targeting CCND2.Objective: The aim of the present study is to obtain studies which are randomized controlled trials applied mesenchymal stem cells(MSCs)in the treatment of osteoarthritis(OA),and to further comprehensively analyze the efficacy and adverse effects of mesenchymal stem cells,and evaluate the feasibility of mesenchymal stem cells in the treatment of osteoarthritis.Methods: We searched several databases including: PubMed,Embase,Scopus,Web of Science,Cochrane Library,Ovid,Science Direct and Springer-Link.The main keywords: "knees","clinical trial","mesenchymal","stem cell" and "osteoarthritis" were retrieved.Inclusion criteria: 1)Randomized controlled test;2)HA(hyaluronic acid,HA)as a control group;3)patients with KOA;4)Patients were injected with mesenchymal stem cells as intervention: Excluded criteria: cell experiments,animal experiments,prospective studies.Results: 870 articles were search in all databases after screening.9 studies were finally included.After 6 to 12 months of mesenchymal stem cell injection,most of scores were significantly improved.Compared with HA group,MSCs could greatly relieve OA symptoms.Meanwhile,we analyzed the adverse effects of MSCs.These include common post-intervention short-term pain,swelling,synovitis,joint effusion and other manifestations.No permanent adverse effects were found except surgical infection and other factors.Conclusion: Mesenchymal stem cells could significantly alleviate the symptoms of osteoarthritis,and mesenchymal stem cells is a prospective treatment in osteoarthritis.