口腔疾病防治 (Aug 2018)

Role of miR⁃27a in the osteogenic differentiation of beagle maxillary sinus membrane stem cells

  • ZHANG Jing,
  • ZHU Shuangxi,
  • RONG Qiong,
  • PENG Wei,
  • LI Xiang,
  • CHEN Songling

DOI
https://doi.org/10.12016/j.issn.2096⁃1456.2018.08.002
Journal volume & issue
Vol. 26, no. 8
pp. 484 – 490

Abstract

Read online

Objective To detect the expression level of miR ⁃ 27a during the osteogenic differentiation of beagle maxillary sinus membrane stem cells (MSMSCs) and explore the role of miR ⁃ 27a in the osteogenic differentiation of MSMSCs. Methods Beagle MSMSCs were cultured in vitro. The expression level of miR⁃ 27a was detected via RT⁃PCR after an osteogenic inductive culture was prepared. The mRNA expression levels of Runx2 and OPN were exam⁃ ined via RT⁃PCR, and the protein expression levels of Runx2 and OPN were examined via Western blot after the cells were transfected with pre⁃miR⁃27a or anti⁃miR⁃27a. Finally, osteoprogenitor cells transfected with pre⁃miR⁃27a were composited with Bio⁃Oss particles and subcutaneously implanted into nude mice to form ectopic bone formation models, and then the inhibition of bone formation from miR⁃27a was observed in vivo. Results The expression level of miR⁃ 27a in the beagle MSMSCs decreased after osteogenic inductive culturing. The relative miR⁃27a levels were significant⁃ ly decreased at day 1 (t＝3.795, P＝0.023), day 3 (t＝4.493, P＝0.011), day 7 (t＝11.591, P < 0.001), day 14 (t＝ 12.542, P < 0.001), and day 21 (t＝5.621, P＝0.008) compared with day 0. In addition, the expression levels of Runx2 mRNA (t＝4.923, P＝0.007) and protein (t＝4.425, P＝0.008) were reduced after the cells were transfected with pre⁃ miR ⁃ 27a. The expression levels OPN mRNA (t＝5.253, P＝0.006) and protein (t＝5.132, P＝0.006) were also re⁃ duced. In contrast, the mRNA expression levels of Runx2 (t＝3.925, P＝0.013) and OPN (t＝3.712, P＝0.019) were in⁃ creased after the cells were transfected with anti⁃miR⁃27a, and bone formation was observed after the subcutaneous im⁃ plantation of beagle MSMSCs composited with Bio⁃Oss in nude mice. Nevertheless, ectopic bone formation was inhibit⁃ ed by pre⁃miR⁃27a⁃transfected beagle MSMSCs composited with Bio⁃Oss (t＝7.219, P＝0.0020). Conclusion MiR⁃ 27a negatively regulates the osteogenic differentiation of MSMSCs.

Keywords