MiR-212 promotes proliferation and inhibits apoptosis of osteosarcoma cells via regulating hedgehog signaling pathway

J BUON. 2020 Jul-Aug;25(4):2086-2091.

Abstract

Purpose: To explore the effects of micro ribonucleic acid 212 (miR-212) on proliferation and apoptosis of osteosarcoma cells via the Hedgehog signaling pathway.

Methods: hFOB1.19 cells were enrolled as normal group, and osteosarcoma MG63 cells were divided into osteosarcoma group and miR-212 mimics group. The cells in the normal and osteosarcoma group were cultured normally, while those in miR-212 mimics group were cultured in medium containing miR-212 mimics. After 24 h of culture, the cells were collected for detection. Quantitative polymerase chain reaction (qPCR) assay was performed to determine the expression level of miR-212. The Gli1 expression level was measured by immunohistochemistry and Western blotting. Additionally, cell proliferation and apoptosis were detected by cell counting kit-8 (CCK-8) assay and flow cytometry, respectively.

Results: Based on immunohistochemistry, the average optical density of Gli1 positive expression was evidently increased in osteosarcoma group and miR-212 mimics group compared with that in normal group. The results of Western blotting and qRT-PCR showed that compared with those in normal group, the relative protein expression level of Gli1 and relative expression level of miR-212 were notably raised in the other two groups, and these levels were remarkably higher in miR-212 mimics group than those in osteosarcoma group. According to CCK-8 assay, the proliferation rate of cells was overtly higher in osteosarcoma group and miR-212 mimics group than that in normal group, and also distinctly higher in miR-212 mimics group than that in osteosarcoma group.

Conclusions: MiR-212 promotes the proliferation and inhibits the apoptosis of osteosarcoma cells by up-regulating the Hedgehog signaling pathway.

MeSH terms

  • Apoptosis / physiology
  • Bone Neoplasms / genetics
  • Bone Neoplasms / metabolism*
  • Bone Neoplasms / pathology
  • Cell Proliferation / physiology
  • Hedgehog Proteins / metabolism*
  • Humans
  • Immunohistochemistry / methods*
  • MicroRNAs / genetics
  • MicroRNAs / metabolism*
  • Osteosarcoma / genetics
  • Osteosarcoma / metabolism*
  • Osteosarcoma / pathology
  • Signal Transduction
  • Transfection

Substances

  • Hedgehog Proteins
  • MIRN212 microRNA, human
  • MicroRNAs