LncRNA HOTAIR Functions As A Competing Endogenous RNA To Inhibit Cell Proliferation By Sponging MiR-130b In Human Nucleus Pulposus Cells

To proficiently master the cultural method of human degenerative NP cells,thereby establishing basis for the successful performance of the latter experiments.Using combined digestion of 0.25% trypsogen and 0.2% type II collagen(Col II),NP cells were isolated and cultured.Col Ⅱ and proteoglycan were found by immunocytochemistry staining and toluidine blue staining.we can successfully culture human degenerative NP cells,and P2 generation of degenerative NP cell is adapt to experiment in vitro.Intervertebral disc degeneration(IDD)is the prominent cause that contributes to low back pain(LBP).Nevertheless,the pathogenesis of IDD is still remain unknown.Differentially expressed lnc RNA HOTAIR were identified using quantitative real-time polymerase chain reaction(q RT-PCR).Next,we investigated the function of HOTAIR in the proliferation of human nucleus pulposus(NP)cells of IDD in vitro and further clarified its mechanism.The expression levels of HOTAIR in NP cells were determined by q RT-PCR.The cell proliferation of NP cells in vitro were investigated by western blot and immunostaining for Proliferative Marker Ki-67 after HOTAIR was overexpression with retroviral vector construction.And the expression levels of cell proliferation related factors were determined to clarify the mechanism.We found that HOTAIR was significantly down-regulated in degenerate NP cells compared with normal NP cells.The proliferation rate was lower in LV-5 HOTAIR NP cells than those in LV-5 NC NP cells.In addition,HTOAIR was confirmed to target mi R-130 b,and mi R-130 b inhibition reversed the proliferation of HOTAIR overexpression on NP cell.In particular,HOTAIR may act as a ce RNA,effectively becoming a sink for mi R-130 b,thereby modulating the derepression of PTEN.PTEN were upregulated signifcantly when HOTAIR was overexpression.While,p-AKT/AKT and Cyclin D1 were downregulated signifcantly when HOTAIR was overexpression.These results indicated that HOTAIR functioned as a protective lnc RNA,which could inhibit the proliferation of dyregulated NP cells in vitro.HOTAIR could be a potential target for the treatment of IDD.