口腔颌面外科杂志2024，Vol.34Issue(3)：163-169,7.DOI:10.12439/kqhm.1005-4979.2024.03.001
LincRNA-EPS对RANKL和LPS诱导RAW264.7细胞破骨分化影响的实验研究
Effect of lincRNA-EPS on osteoclastogenesis of RAW264.7 cells induced by RANKL and LPS:An experimental study
摘要
Abstract
Objective:To study the effect of long non-coding RNA-EPS(lincRNA-EPS)on osteoclastogenesis of RAW264.7 cells induced by receptor activator for nuclear factor-κB ligand(RANKL)and lipopolysaccharide(LPS).Methods:RAW264.7 cell strain overexpressing lincRNA-EPS(lincRNA-EPS OE strand)and its negative control strain were generated by transfection of lincRNA-EPS overexpressing vector and negative control vector and subsequent screening.Osteogenesis was induced with either RANKL or LPS(after pretreatment with RANKL).The cells were then stained with tartrate resistant acid phosphatase staining(TRAP staining)kit to count the number and observe the morphology of osteoclasts.Phalloidin staining was performed to detect the F-actin rings and therefore to evaluate bone-resorbing ability of osteoclasts.Real-time quantitative polymerase chain reaction(RT-qPCR)was performed to measure the mRNA levels of osteoclast-associated genes.Results:RAW264.7 cell strain overexpressing lincRNA-EPS and its negative control strain were generated.Neither strain showed significant difference during osteoclastogenesis(P>0.05),either induced by RANKL or LPS;for osteoclastogenesis by LPS induction,TRAP staining showed that significantly more osteoclasts were generated from lincRNA-EPS OE strain(P<0.05),while phalloidin staining showed that larger F-actin rings were generated from lincRNA-EPS OE strain,and RT-qPCR also showed that the mRNA levels of 4 osteoclastogenesis-related genes(TRAP,CTSK,DC-STAMP and ATP6v0d2)after induction were significantly lower in lincRNA-EPS OE strain(P<0.05);however,those difference were not present(P>0.05)when osteoclastogenesis was induced by RANKL.Conclusion:The overexpression of lincRNA-EPS inhibits LPS-induced osteoclastogenesis and the function of osteoclasts.However,the overexpression of lincRNA-EPS does not affect RANKL-induced osteoclastogenesis significantly.关键词
长链非编码RNA/牙周炎/破骨分化/脂多糖Key words
long non-coding RNA/periodontitis/osteoclastogenesis/lipopolysaccharide分类
医药卫生引用本文复制引用
张栩,王雅冰,苏俭生..LincRNA-EPS对RANKL和LPS诱导RAW264.7细胞破骨分化影响的实验研究[J].口腔颌面外科杂志,2024,34(3):163-169,7.基金项目
国家自然科学基金(82170913、82201077) （82170913、82201077）
上海市科学技术委员会项目(201409006200) （201409006200）
