Long non-coding RNA DANCR enhances chondrogenic differentiation and proliferation of human synovium-derived mesenchymal stem cells / 中国组织工程研究

ABSTRACT

BACKGROUND: A number of studies have shown that long non-coding RNA DANCR can play an important role in various pathophysiological processes through Wnt/β-catenin signaling pathway.OBJECTIVE: To explore the effect of long non-coding RNA DANCR on the proliferation and chondrogenesis of synovium-derived mesenchymal stem cells.METHODS: Passage 3 synovium-derived mesenchymal stem cells were obtained and transfected with pcDNA3.1-GP (control) and pcDNA3.1-GP(DANCR Homo) (experimental). Cell viability was estimated at 1, 2, 3, 4, 5, 6 and 7 days after DANCR transfection. The passage 3 cells were cultured in the chondriogenic medium for 14 days. And the chondrogenesis potential of cells was examined by toluidine blue staining. The chondrogenic-specific marker genes Aggrecan, Type II collagen (Col2) and Sox9 were determined by Real-time PCR.RESULTS AND CONCLUSION: The synovium-derived mesenchymal stem cells exhibited S-shaped curves in the two groups, with cell arrest at 1-2 days and rapid proliferation beginning at 3 days. Cell counting kit-8 assay and toluidine blue staining showed overexpressing DANCR significantly promoted proliferation in synovium-derived mesenchymal stem cells. The aggregates from synovium-derived mesenchymal stem cells in the experimental group had a greater amount of toluidine blue staining than the control group. In addition, we detected the higher expression of chondrogenic specific marker genes, such as Col2, Sox9 and Aggrecan, in the experimental group than the control group at 14 days after chondrogenic induction (P < 0.05). These results demonstrate that long non-coding RNA DANCR could enhance chondrogenic differentiation and proliferation of human synovium-derived mesenchymal stem cells and increase the expression of chondrogenic specific marker genes.