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Tumor Biology

, Volume 37, Issue 10, pp 13469–13477 | Cite as

MicroRNA-106b promotes pituitary tumor cell proliferation and invasion through PI3K/AKT signaling pathway by targeting PTEN

  • Kai Zhou
  • Tingrong Zhang
  • YanDong Fan
  • Serick
  • Guojia Du
  • Pengfei Wu
  • Dangmurenjiafu Geng
Original Article

Abstract

The purpose of this study was to investigate the expression of microRNA-106b (miR-106b) and phosphatase and tensin homolog deleted on chromosome 10 (PTEN) in pituitary tumor and to confirm whether miR-106b promotes proliferation and invasion of pituitary tumor cells through the PI3K/AKT signaling pathway by targeted regulation of PTEN expression, and thereby to find new targets for the treatment of pituitary tumor. Fifty-five cases of pituitary tumor tissue samples were collected, including 29 cases of invasive pituitary tumor, non-invasive 26 cases, and 8 normal pituitaries. The expression level of miR-106b in pituitary tumor tissue was detected by quantitative real-time PCR, and the expression of PTEN protein was detected by immunohistochemistry. PTEN 3′-untranslated region (UTR) luciferase vector was constructed, and dual-luciferase reporter gene assay was employed to examine the effect of miR-106b on PTEN 3′-UTR luciferase activity. AtT-20 cells were transfected with miR-106b mimics, miR-106b inhibitor, PTEN expression plasmid, and miR-106b mimics + PTEN expression plasmid respectively, and the changes in cellular proliferation and invasion were observed via MTT method and transwell assay respectively. PTEN messenger RNA (mRNA) expression was determined by quantitative real-time PCR, and western blotting was performed to detect the expression of PTEN, PI3K, AKT, and pAKT. miR-106b showed up-regulation in invasive pituitary tumor tissue: the expression level was significantly up-regulated compared with normal tissues and the non-invasive pituitary tumor tissue (P < 0.05). The positive rate of PTEN protein expression in invasive pituitary tumor tissues was significantly lower than in normal and non-invasive tissues (P < 0.01). Dual-luciferase reporter gene assay showed that miR-106b could bind to the 3ʹ-UTR of PTEN specifically and significantly inhibited the luciferase activity, cutting the 46 % (P < 0.01). Down-regulation of miR-106b or up-regulation of PTEN could suppress cell proliferation and invasion of AtT-20 cells, and PTEN expression plasmid could partially simulate the function of miR-106b. Expression of PTEN mRNA and protein decreased significantly in AtT-20 cells overexpressing miR-106b. The expression levels of PI3K and p-AKT were significantly inhibited by miR-106b inhibitor and increased by miR-106b mimics. The expression of miR-106b showed up-regulation in pituitary tumor tissues, while the protein expression of PTEN presented opposite results. The findings of this study further demonstrated that miR-106b as an oncogene regulated the pituitary tumor cell proliferation and invasion in vitro by directly targeting PTEN through the PI3K/AKT signaling pathway. Our study suggests that miR-106b and PTEN are likely to serve as potential diagnostic biomarkers or therapeutic targets for pituitary tumor treatment in the future.

Keywords

Pituitary tumor miR-106b PTEN PI3K/AKT Cell proliferation Invasion AtT-20 cells 

Notes

Acknowledgments

This work was supported by Natural Science Foundation of Xinjiang Uygur Autonomous Region (2013211A078).

Compliance with ethical standards

Informed consent for this study was obtained from all patients, and this study was approved by the Research Ethics Committee of the First Affiliated Hospital of Xinjiang Medical University.

Conflicts of interest

None

Supplementary material

13277_2016_5155_MOESM1_ESM.jpg (585 kb)
Supplementary Figure 1 The mechanistic schema associating miRNA-106b/PTEN/PIK3/AKT with proliferation and invasion of pituitary tumor cells. PTEN expressions are inhibited by targeting of miR-106b to 3′ UTR of PTEN mRNA, thereby deterring the inhibitory effects of PTEN on conversion of PIP3 to PIP2. Accumulating PIP3 would then phosphorylate AKT after its combination with PDK1 and PDK2. Subsequently, phosphorylated AKT would translocate to the nucleus, activating molecules relevant to cell migration (e.g. MMP-3, MMP9 and VEGF) and cell proliferation (e.g. GSK3β). PIP3, phosphatidylinositol 3,4,5-triphosphate; PIP2, phosphatidylinositol 4,5-bisphosphate; PDK, phosphoinositide-dependent kinase; MMP, matrix metallo proteinase; VEGF, vascular endothelial growth factor. (JPEG 584 kb)

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Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2016

Authors and Affiliations

  • Kai Zhou
    • 1
  • Tingrong Zhang
    • 1
  • YanDong Fan
    • 1
  • Serick
    • 1
  • Guojia Du
    • 1
  • Pengfei Wu
    • 1
  • Dangmurenjiafu Geng
    • 1
  1. 1.Department of Neurosurgerythe First Affiliated Hospital of Xinjiang Medical UniversityUrumqiChina

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