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. 2018 Aug 14:11:4893-4899.
doi: 10.2147/OTT.S170293. eCollection 2018.

Downregulation of lncRNA ANRIL suppresses growth and metastasis in human osteosarcoma cells

Hongya Guan  1 , Yingwu Mei  2 , Yang Mi  2 , Cheng Li  3 , Xiaoya Sun  2 , Xuefeng Zhao  2 , Jia Liu  1 , Wei Cao  1 , Yuebai Li  2 , Yisheng Wang  3
Affiliations

Downregulation of lncRNA ANRIL suppresses growth and metastasis in human osteosarcoma cells

Hongya Guan et al. Onco Targets Ther. .

Retraction in

Abstract

Background: This study was designed to research the potential function of lncRNA ANRIL in osteosarcoma (OS).

Materials and methods: Quantitative real-time PCR, cell counting kit-8, wound healing assay, Transwell assay, flow cytometric analysis, caspase activity analysis, and Western blot were carried out.

Results: ANRIL was remarkably upregulated in human OS tissues and cells, and knockdown of ANRIL significantly suppressed MG63 cell proliferation, migration, and invasion and promoted apoptosis. Moreover, our mechanistic research findings verified that ANRIL-influenced growth and apoptosis may be partly through regulation of caspase-3 and Bcl-2. Migration and invasion were influenced via ANRIL-mediated regulation of MTA1, TIMP-2, and E-cadherin.

Conclusion: Our finding demonstrates that ANRIL plays vital roles in OS growth and metastasis.

Keywords: ANRIL; apoptosis; invasion; long noncoding RNA; osteosarcoma; proliferation.

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Conflict of interest statement

Disclosure The authors report no conflicts of interest in this work.

Figures

Figure 1
Figure 1
lncRNA ANRIL expression in OS tissues and cell lines. Notes: (A) lncRNA ANRIL expression level in 30 OS tissues compared with adjacent nontumor tissues. (B) lncRNA ANRIL expression level in 4 OS cell lines (HOS, U-2OS, MG63, and SAOS-2) and normal human osteoblast hFOB1.19 cell line was determined by qRT-PCR. The values are given as mean ± SD of 3 independent experiments. **P<0.01. Abbreviations: lncRNA, long noncoding RNA; OS, osteosarcoma; qRT-PCR, quantitative real-time PCR.
Figure 2
Figure 2
(A) qRT-PCR displayed that ANRIL expression significantly decreased in MG63 cells after treatment with si-ANRIL. (B) CCK-8 assay revealed that knockdown of ANRIL could significantly repress MG63 cell proliferation. **P<0.01. Abbreviations: CCK-8, cell counting kit-8; lncRNA, long noncoding RNA; NC, negative control; qRT-PCR, quantitative real-time PCR.
Figure 3
Figure 3
(A–C) Flow cytometry and caspase-3 activity assay showed that ANRIL knockdown in MG63 cells significantly increased cell apoptosis ability. (D, E) Cells with ANRIL knocked down exhibited repressed expression of Bcl-2 protein and elevated expression of caspase-3 protein. **P<0.01. Abbreviation: NC, negative control.
Figure 3
Figure 3
(A–C) Flow cytometry and caspase-3 activity assay showed that ANRIL knockdown in MG63 cells significantly increased cell apoptosis ability. (D, E) Cells with ANRIL knocked down exhibited repressed expression of Bcl-2 protein and elevated expression of caspase-3 protein. **P<0.01. Abbreviation: NC, negative control.
Figure 4
Figure 4
(A) Wound healing assay showed that downregulating ANRIL expression dramatically decreased MG63 cell migration. (B, C) Transwell assay showed that suppressing ANRIL expression reduced MG63 cell invasiveness. (D, E) Cells with ANRIL knocked down exhibited increased TIMP-2 and E-cadherin protein expression and decreased MTA1 protein expression. *P<0.05, **P<0.01. The Original magnification, ×100. Scale bars= 200 μm. Abbreviation: NC, negative control.
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